DNA forms not only the canonical duplex but also various non-canonical structures such as triplex, G-quadruplex, and i-motif. The are many external factors that influence folding and stability of the individual forms. Further, DNA structure can be affected by attachment of various artificial covalent or noncovalent ligands.

Our investigations are focused on detailed structural characterization of short purine oligonucleotides clipped by proper sequential blocks. For this purpose, modern NMR experiments combined with MD simulations are employed. The effect of modification of selected nucleotide on the structural properties of designed models is characterized to gain deeper understanding of key noncovalent interactions that contribute to the DNA folding.

Examples of PhD topics:
a) Structure of parallel forms of nucleic acids studied by NMR spectroscopy and molecular modelling
b) Designing modified DNA fragments

More information:
radek.marek@ceitec.muni.cz
jan.novotny@ceitec.muni.cz

Notes

Note: All candidates should contact R. Marek for informal discussion before initiating the formal application process.

Supervisor

prof. RNDr. Radek Marek, Ph.D.

V současné době máme k dispozici nadkritické množství informací ohledně proteinových strukturních rodin. Konkrétně, pro většinu rodin známe stovky struktur jejích zástupců, přičemž tyto struktury pocházejí z různých organismů, některé z nich váží rozličné ligandy a mnohé obsahují různorodé mutace. Tyto informace umožňují analýzu „anatomie“ daných proteinových rodin. Například studium elementů sekundární struktury (šroubovic a skládaných listů), jejich vzájemného uspořádání, konzervovanosti a určování, které z těchto elementů jsou pro danou proteinovou rodinu klíčové a které se vyskytují jen raritně. Dále pak zkoumání proteinových tunelů a pórů, jejich charakteristik a četnosti jejich výskytu u jednotlivých zástupců proteinové rodiny. V rámci laboratoře LCC jsou vyvíjeny softwarové nástroje pro realizaci výše uvedených analýz, např. software MOLE, LiteMol, SecStrAnalyzer. Hlavním cílem disertační práce je zaměřit se na několik konkrétních biologicky významných proteinových rodin (např. cytochromy, poriny, dehalogenázy, proapoptotické proteiny) a provést jejich detailní analýzu. Dalším cílem je spolupráce při vývoji uvedených softwarových nástrojů.

Notes

Vypsáno pro přihlášení studentky Jany Porubské.

Supervisor

prof. RNDr. Radka Svobodová, Ph.D.

Correlative Light Electron Microscopy (CLEM) uses a combination of an optical (fluorescence) microscope and a cryo-electron microscope. Two images of the sample are taken simultaneously – one with the optical light, the other with the electron beam. This technology allows to capture not only dynamic changes but also the molecular ultrastructure of living systems. New developments in accurate positional referencing of specimens on mounting grids, advances in the instrumentation, and the availability of software packages for cross-platform data correlation allow to image the ultrastructure of nucleolar sub-compartments and to track specific proteins found in phase-separated organelles. In this project, we will implement the CLEM technology to investigate and visualize phase-separated organelles involved in transcription by RNA polymerase II and investigate their regulatory mechanism during transcription. This biophysically focused project will also involve other imaging approaches, including single-particle reconstruction cryo-electron microscopy and cryo-electron tomography, which will help to obtain an overall picture of condensate-based transcription at different resolutions.

Supervisor

prof. Mgr. Richard Štefl, Ph.D.

Objective

The main objective of this PhD project is the simulation-based design of new peptidoglycan (PG)-binding motifs and peptides with tailored affinity and selectivity toward Gram-positive bacterial cell walls. The project will identify the molecular features and structural patterns that enable selective recognition of bacterial PG and use these principles for the de novo design of new PG binders through a combination of molecular simulations and genetic algorithms. Selected sequences will subsequently be prioritized for experimental verification. The resulting binders could provide versatile molecular scaffolds for the development of more selective antibacterial strategies and contribute to next-generation approaches for targeting drug-resistant bacterial infections.

Description

Bacterial drug resistance is a growing public health threat that demands new therapeutic approaches. One promising direction involves endolysins - enzymes produced by bacteriophages, the viruses that infect bacteria. Endolysins degrade PG, the cell-wall macromolecule that protects bacterial cells, and can therefore kill bacteria in a highly targeted manner. However, their therapeutic application is constrained by stability, delivery, and fragmentary molecular understanding of how their cell wall-binding domains (CBDs) recognize PG, hampering rational optimization without compromising bacterial targeting. This PhD project will use biophysical and biochemical methods to investigate how endolysin CBDs and their molecular motifs recognise PG architectures in Gram-positive bacteria, including drug-resistant strains. These CBDs will serve as reference systems for identifying the molecular features that drive selective PG recognition. The resulting principles will then be translated into the design of new, smaller, and tunable PG-binding sequences with tailored affinity and selectivity. The student(s) will gain training in high-performance computing, molecular modeling of complex biomolecular systems, atomistic molecular dynamics simulations, free energy methods, and genetic algorithms. Machine-learning approaches may also be incorporated to accelerate the exploration of sequence space. Overall, the project is expected to advance the molecular understanding of PG recognition and support the development of computationally designed binders for more selective antibacterial strategies.

Additional information

Before the formal application process, all interested candidates should contact Dr. Denys Biriukov (denys.biriukov@ceitec.muni.cz).
More about the supervisor and his team.

Supervisor

Denys Biriukov, Ph.D.

Současné chemické, biochemické i biologické experimentálních metody produkují mnoho různorodých výstupních dat. Tato data jsou nejen důležitou součástí publikací, ale i velmi cenným materiálem k dalším analýzám. Proto v rámci grantových projektů i přímo mezi výzkumnými pracovníky silně vyvstává potřeba strukturovaného zaznamenávání výstupních dat a postupů, realizovaných v průběhu experimentů.  

K tomuto účelu začala být v posledních letech intenzivně vyvíjena softwarová řešení – takzvané elektronické laboratorní deníky (ELN). Vývoj těchto softwarových nástrojů v současné době dynamicky probíhá a je dostupno několik desítek open source i komerčních řešení. Pro výzkumné pracovníky je velkou výzvou zjistit, který z těchto nástrojů je vhodný pro jejich laboratoř a integrovat jej do datového workflow své laboratoře. A právě na tuto oblast je cílena disertační práce. 

Konkrétní úkoly práce jsou: Zorientovat se v pluralitě ELN, analyzovat současnou situaci a vyhledat vhodná řešení pro vybrané typy laboratoří. Integrovat ELN do bioinformatických datových workflow vybraných laboratoří a případně provést adaptace a rozšíření ELN, nutné pro jejich efektivní využívání.

Notes

The topic is reserved.

Supervisor

prof. RNDr. Radka Svobodová, Ph.D.

We invite enthusiastic application for a PhD position with interest in molecular biology and biochemistry. The successful candidate will work under the supervision of Dr. Krejčí to identify and characterise novel inhibitors of DNA repair nucleases, their mechanisms of action and therapeutic implications.

The PhD position candidate should hold or be about to complete a Masters degree in molecular biology, biochemistry or similar field. The applicant is also expected to demonstrate essential training in a range of molecular biology techniques relevant to basic research, should be well-organised, motivated and passionate about pursuing a career in biomedical research.

We offer fully funded positions with competitive salary in a well established laboratory. The lab hosts international team members, has a strong publication track record and international collaborations. The offered projects contribute to a rapidly advancing, very competitive field. The successful candidate can start immediately.

Supervisor

doc. Mgr. Lumír Krejčí, Ph.D.

OBJECTIVES: The aim is to elucidate the relationship between molecular properties of amphiphilic peptides and their ability to translocate and form transmembrane pores in membranes with various lipid compositions. The obtained understanding will be used for the development of new antimicrobial peptides, which can serve as a new type of antibiotic drugs.



DESCRIPTION: Antibiotic-resistant bacteria cause more than 700 000 deaths per year, and the forecast is 10 million per year in 2050. Moreover, emerging strains of bacteria resistant to all available antibiotics may lead to a global post-antibiotic era. Because of this threat, the WHO and the UN are encouraging the research and development of new treatments. Antimicrobial peptides are promising candidates for such new treatments. We will study the molecular mechanism of action of antimicrobial peptides and determine the critical peptide properties required for membrane disruption via the formation of transmembrane pores and spontaneous peptide translocation across membranes. Based on the obtained insight, we will design new peptides and test their abilities. The most effective peptides will be evaluated for antimicrobial activity and human cell toxicity using growth inhibition and hemolytic assays, respectively. Student(s) will master tools of computer simulations, in particular, molecular dynamics techniques and methods to calculate free energies. Moreover, he/she will learn the advantages and disadvantages of various protein and membrane parameterizations, including all-atom and coarse-grained models. The simulations will be complemented by in vitro experiments using fluorescent techniques.



EXAMPLES of potential projects: * Antimicrobial peptides and formation of membrane pores * Synergistic mechanisms between antimicrobial peptides * Membrane disruption by antimicrobial peptides in non-equilibrium conditions



MORE INFORMATION about the group: vacha.ceitec.cz



PLEASE NOTE: before the formal application process, all interested candidates should contact Robert Vacha (robert.vacha@mail.muni.cz).

Supervisor

prof. RNDr. Robert Vácha, PhD.

Peptidová/proteinová afinita k membránám je závislá na konkrétní sekvenci a membránovém složení. Bohužel porozumění tohoto komplexního vztahu nám dosud chybí. Cílem tohoto projektu odhalit tento vztah a využít ho k vývoji nových antimikrobiálních peptidů, biomarkerů a senzorů.

Student získá znalosti v oblasti fluorescence, lipidových váčků, QCM.

Supervisor

prof. RNDr. Robert Vácha, PhD.

OBJECTIVES: The aim is to elucidate the relationship between protein sequence and preferred composition and curvature of human membranes,i.e., find peptide motifs that are selective to specific membranes in cells (plasma membrane, endoplasmic reticulum, Golgi apparatus, mitochondria, etc.). The obtained understanding will be used for the development of new protein biomarkers, sensors, scaffolds, and drugs.



DESCRIPTION: The control of biological membrane shape and composition is vital to eukaryotic life. Despite a continuous exchange of material, organelles maintain a precise combination and organization of membrane lipids, which is crucial for their function and the recruitment of many peripheral proteins. Membrane shape thus enables the cell to organize proteins and their functions in space and time, without which serious diseases can occur. Moreover, membrane curvature and lipid content can be specific to cancer cells, bacteria, and enveloped virus coatings, which could be utilized for selective targeting. We will develop a new method, using which we will elucidate the relationship between the protein sequence and the preferred membrane. The relationship will lay the foundations for the design of new protein motifs sensitive to membranes with a specific curvature and composition. Student(s) will master tools of computer simulations, in particular, molecular dynamics techniques and methods to calculate free energies. Moreover, he/she will learn the advantages and disadvantages of various protein and membrane parameterizations, including all-atom and coarse-grained models.



EXAMPLES of potential projects: * Determination of helical motifs for specific membrane compositions * Development of implicit membrane model for fast determination of protein-membrane affinity * Helical peptides and their sensitivity for membrane curvature



MORE INFORMATION: vacha.ceitec.cz



PLEASE NOTE: before the formal application process, all interested candidates should contact Robert Vacha (robert.vacha@mail.muni.cz).

Supervisor

prof. RNDr. Robert Vácha, PhD.

The research goal is investigation of structure, dynamics, and biologically relevant properties of proteins, using NMR spectroscopy and other high-resolution approaches. Currently, our group is mostly interested in studies of molecular motions using NMR relaxation and relaxation dispersion; in studies of protein disorder using NMR approaches providing sufficient resolution (usually based on non-uniformly sampled high-dimensional spectra); and in studies of interactions of intrinsically disordered proteins with their binding partners (using NMR, cryo-EM, and biophysical methods). The systems currently studied in the laboratory include bacterial RNA polymerases and microtubule associated proteins.

We are inetrested structure and dynamics of well-ordered and domains of subunits and sigma factors of RNA polymerase from B. subtilis, characterization of structural features and dynamics of disordered domain, and in importance of electrostatic interactions for structural properties and biological function of the protein. Currently we extend our interest to mycobacterial RNA polymerase.

Microtubule associated protein 2c (MAP2c) is a key factor regulating microtubule dynamics in developing brain neurons, and an example of an intrinsically disordered proteins with an important physiological function and detectable structure-function relationship. The first goal is to study MAP2c in a natural complexity and by methods providing atomic resolution. Such methods include paramagnetic relaxation interference, to detect and describe transient local structures of MAP2c important for its function, and real-time NMR, to monitor kinetics of MAP2c phosphorylation by relevant kinases of different signalling pathways. The second goal is to characterize interactions of MAP2c with biologically important binding partners, especially with isoforms and a monomeric form of regulatory protein 14-3-3. The third goal is to test the effect of cellular environment on MAP2c by recording NMR spectra at near-to-native conditions (in cells and/or cell lysates) and/or by performing cryo-electron tomography on monolayered neurons.

EXAMPLES OF POTENTIAL PHD TOPICS:

• Interactions underlying physiological function of Microtubule Associated Protein 2c
• Structure, dynamics and interactions of bacterial RNA polymerase subunits and sigma factors

Supervisor

prof. Mgr. Lukáš Žídek, Ph.D.

BACKGROUND: Several neurodegenerative diseases are associated with the formation of fibrous protein aggregates. The fibrillization of amyloid beta peptide into amyloid plaques and the agregation of hyperphosphorylated tau protein into neurofibrillar tangles are main neuropatological signs of Alzheimer disease. Studying of how different factors influence the formation of biomolecular complexes is the key for understanding underlying molecular mechanism of neurodegerative processes. The described activities are part of international research projects allowing to spend the part of PhD study at the collaborative groups in Europe or North and South America and to learn specific research techniques, there.

OBJECTIVES: The research aims to elucidate molecular mechanisms of conformational changes leading to the modified potential of biomolecular complex formation. Interdisciplinary approach combining computational biophysical chemistry, structural biology, bioinformatics and biophysical interaction techniques will be applied.

FOCUS: Doctoral research projects focus on the monitoring of post-translational modification of studied proteins, their interaction with adaptor proteins and induced conformational changes. Students benefit from outstanding research facilities of CEITEC-MU that include cryoEM tomography, NMR, AFM, and biophysical interaction methods.

EXAMPLES of potential student doctoral projects:

• Are Tau fibrils induced by phosphorylation and the interaction with 14-3-3 proteins relevant for Alzheimer disease?
• A Tau conformational changes induced by phosphorylation and 14-3-3 proteins relevant in neurodegenerative diseases
• Oligomerization states within the 14-3-3 protein family
• Computational prediction of biomolecular complexes and their statibities

MORE INFORMATION: jozef.hritz@ceitec.muni.cz

PLEASE NOTE: before initiating the formal application process to doctoral studies, all interested candidates are required to contact Jozef Hritz (jozef.hritz@ceitec.muni.cz) for informal discussion.

Supervisor

doc. RNDr. Mgr. Jozef Hritz, Ph.D.

We apply structural biology methods in order to gain a mechanistic view of CK1ε action in the Wnt signalling pathways. CK1ε represents an attractive therapeutic target but currently two key steps in the CK1ε-mediated Wnt signal transduction are unclear: how CK1ε gets activated and/or engages target proteins in response to Wnt signal and how CK1ε phosphorylates its key substrate Dishevelled (DVL).

Our preliminary data suggest that we can efficiently apply methods of integrated structural biology to (i) probe the DVL conformational landscape using in vitro and in vivo FRET sensors coupled to SAXS and CryoEM, (ii) understand the (auto)phosphorylation regulatory mechanisms of CK1ε, (iii) analyse by NMR the functional consequences of DVL phosphorylation and (iv) monitor DVL phosphorylation by real-time NMR under controlled cellular conditions. The position is part of a multidisciplinary project that combines (i) cellular and molecular biology, (ii) proteomic analysis, (iii) biochemistry and structural biology, and received generous funding in a very competitive grant scheme.

Keywords: CK1ε, WNT, DVL phosphorylation, SAXS, cryo-EM, cryo-electron microscopy, real-time NMR

Contact:
Kostas Tripsianes, PhD | CEITEC - Central European Institute of Technology | Masaryk University | Kamenice 5/A35/1S081, CZ-62500 Brno | phone: 00420 549 49 6607

Supervisor

Konstantinos Tripsianes, Ph.D.

Our scientific goal is understanding of the most basic principles of structural dynamics, function and evolution of DNA and RNA.

To achieve our goal, we use a wide portfolio of theoretical/computational approaches. Our research is closely related to experiments, mostly via extensive collaborations, though in the prebiotic chemistry we have in house experiments. We offer thesis essentially on any topic that is currently active in the laboratory. You can get the most up-to-date idea about our current research from the WOS or SCOPUS databases, where you can find all our publications (Sponer, J.), see all our collaborators, etc. The laboratory is located at the Institute of Biophysics, Czech Academy of Sciences, Kralovopolska 135, Brno, where we have a powerfull and regularly upgraded set of high-perfomance computer clusters dedicated exclusively to our group

Our methods are:

• Classical Molecular Dynamics (MD) simulations. Besides standard simulations, we have years of experience in using all classes of enhanced-sampling techniques. We play also a prominent role in development of DNA/RNA simulation force fields and our versions are used world-wide
• Quantum-chemical (QM) method. We are using a wide spectrum of methods, ranging from ultra-accurate computations of small model systems, through large-scale QM studies on biomolecular building blocks with hundreds of atoms up to sophisticated methods that are used in studies of excited states and photochemistry; the later technique is especially relevant to study the origin of life chemistry under UV light. Again, please see the papers we have published in last years.
• Hybrid quantum-classical (QM/MM) methods, quantum molecular dynamics
• Structural bioinformatics

Specific experiments are possible in the field of prebiotic chemistry in collaborating laboratories. Modern computations are extensively combined with many experimental techniques (NMR, X-Ray, high-energy lasers, biochemical techniques) mostly via numerous collaborations. We collaborate with 30 foreign and Czech laboratories. We publish about 20 papers annually and belong to the most cited Czech research groups. We currently work in several mutually interrelated research areas.

• RNA structural dynamics, folding and catalysis
• Protein-RNA (or DNA) complexes. We try to go beyond the ensemble-averaged picture of experimental methods in order to understand how rarely accessed dynamical conformations invisible to experiments allow to separate affinity for reactivity or selectivity.
• DNA, with focus on G-quadruplexes, specifically advanced studies of quadruplex folding mechanisms
• Diverse types of quantum-chemical studies on nucleic acids systems
• Origin of life (prebiotic chemistry), i.e., creation of the simplest chemical life on our planet (or anywhere else in the Universe), with a specific attention paid to the formamide pathway to template-free synthesis of the first RNA molecules. This specific project includes also in house experimental research.

Besides studies of specific systems, we are also involved extensively in method testing/development, mainly in the field of parametrization of molecular mechanical force fields for DNA

NOTE: before initiating the formal application process to doctoral studies, all interested candidates are required to contact Prof. Jiri Sponer (sponer@ncbr.muni.cz) for an informal discussion.

Laboratory web page https://www.ibp.cz/en/research/departments/structure-and-dynamics-of-nucleic-acids/info-about-the-department

List of publications https://www.ibp.cz/en/research/departments/structure-and-dynamics-of-nucleic-acids/publications

Supervisor

prof. RNDr. Jiří Šponer, DrSc.

This PhD project will focus on the development of new molecular inhibitors of Argonaute proteins. Using structure-guided computational design, the student will generate peptide or protein binders targeting key functional surfaces of Argonaute proteins involved in small-RNA loading and formation of the RNA-induced silencing complex (RISC). Selected candidates will be experimentally validated for binding affinity, specificity, and their ability to perturb Argonaute conformational states. Their functional effects will be tested in biochemical loading assays, including guide loading, strand selection, and RISC maturation. The most promising inhibitors will be further analyzed in cell-based assays to determine their impact on Argonaute-bound small RNAs and downstream RNA-silencing activity. This work will be carried out in close collaboration with colleagues at the University of Vienna, who will contribute advanced cellular and RNA-biology assays. The project aims to establish new molecular tools for controlled modulation of Argonaute function and for dissecting the mechanisms of RNA silencing.

Supervisor

prof. Mgr. Richard Štefl, Ph.D.

Our laboratory is focusing on study of molecular mechanisms of genome instability associated diseases
linked to DNA repair defects. DNA in cells is constantly damaged not only from external but also internal sources resulting in accumulation of hundreds of thousand lesion per cell and day. One of the mechanisms involved in genome stability is homologous recombination and its defects are linked to development of various cancers and diseases (BLM, RTS, FA, etc.).

PhD project might involved following topics: 1)RecQ4 helicase, mutated in „Rothmund-Thomson Syndrome“, a its biochemical and biological characterisation; 2) Development of new nuclease inhibitors and their preclinical characterisation; 3) Rad51 paralogs and their role in genome stability and cancer development; 4) Role of G4 structures and their metabolism in genome stability.

Our approaches involve broad range of molecular-biological, biochemical, biophysical, cell biological, genetic and structural methods.

Supervisor

doc. Mgr. Lumír Krejčí, Ph.D.

Supervisor

prof. RNDr. Radek Marek, Ph.D.

V současné době produkují pokročilé experimentální a výpočetní metody ve strukturní biologii rozsáhlé množství dat. Jedná se například o mapy elektronové hustoty z kryoelektronové mikroskopie, buněčné tomogramy, obrazová data ze světelné mikroskopie nebo trajektorie z molekulově-dynamických simulací. Tato data obsahují cenné informace o struktuře a dynamickém chování biomakromolekul v čase. Vzhledem k jejich velikosti a komplexnosti je však pro vědeckou komunitu často obtížné s nimi efektivně pracovat. Bez odpovídajících vizualizačních přístupů se stávají nepřehlednými, což výzkumníkům ztěžuje jejich správnou interpretaci a pochopení širšího biologického kontextu studovaných systémů.

Aby bylo možné využít potenciál těchto datových sad, je nezbytné existující vizualizační nástroje dále rozvíjet a rozšiřovat tak, aby umožňovaly jejich intuitivní zkoumání. Tato disertační práce se proto zaměří na další vývoj metod pro vizuální analýzu biomakromolekul v rámci ekosystému Mol*. Výzkum bude řešit tři klíčové výzvy: efektivní reprezentaci rozsáhlých volumetrických a dynamických dat, plynulou integraci různorodých strukturních dat s funkčními anotacemi a návrh interaktivních nástrojů pro snadné vyhledávání a zobrazení klíčových strukturních prvků, jako jsou například tunely a vazebná místa. Výsledkem tak bude ucelená sada metod, která vědcům poskytne intuitivní prostředí pro studium rozmanitých molekulárních systémů a pochopení jejich strukturně-funkčních vztahů.

Supervisor

RNDr. Tomáš Raček, Ph.D.

Glykoproteomika je nově vznikající obor, který odhaluje souvislosti glykoforem proteinů s rozvojem onemocnění. V organismu je až 80% všech proteinů posttranslačně modifkovaných glykosylací ovlivňující mnoho biologických procesů. Struktura glykanů a místa glykosylace na proteinu mohou být různá, čímž vznikají proteoformy s různými funkcemi, které mohou aktivovat nebo inhibovat různé buněčné procesy. Oblast glykoproteomiky tedy odhaluje tyto složité vztahy, jejich souvislosti se zdravím a nemocemi a je tedy využívána pro identifikaci dalších biomarkerů v oblasti diagnostiky nejen onkologických onemocnění.
Cílem této práce bude využití klinického materiálu (sér pacientů s definovaným onkologickým onemocněních a zdravých dárců) pro identifikaci a label-free kvantifikaci změněných glykoforem vázaných na proteinech. K tomu budou využívané proteomické analýzy založené na měření hmotnostním spektrometrem Fusion Orbitrap (Thermo Fisher Scientific). Data budou hodnocena proteomickými programy (Byonic, Peaks, Proteome Discoverer) a pomocí pokročilých statistických nástrojů v programovacím jazyku R budou definované peptidy se specifickými glykany, které jsou významné pro danou skupinu pacientů. Následně bude pomocí strojového učení hodnoceno, zda dané glykoformy peptidů pomohou zlepšit diagnostiku nebo postup v léčby onemocnění.
Doporučená literatura: (1) Fang, K., Long, Q., Liao, Z. et al. Glycoproteomics revealed novel N-glycosylation biomarkers for early diagnosis of lung adenocarcinoma cancers. Clin Proteom 19, 43 (2022). https://doi.org/10.1186/s12014-022-09376-8, (2) Kim EH, Misek DE. Glycoproteomics-based identification of cancer biomarkers. Int J Proteomics. 2011; 2011:601937. https://doi.org/10.1155/2011/601937, (3) Pan, J., Hu, Y., Sun, S. et al. Glycoproteomics-based signatures for tumor subtyping and clinical outcome prediction of high-grade serous ovarian cancer. Nat Commun 11, 6139 (2020).
https://doi.org/10.1038/s41467-020-19976-3

Supervisor

prof. Ing. Lenka Hernychová, Ph.D.

Detection of tumor biomarkers is essential for early diagnostics of cancer, since it helps to decrease mortality and high cost associated with late treatment, and is also highly beneficial when monitoring response to therapy or possibility of relapse. In recent years, various analytical methods based on electrochemical (EC) detection have been reported. These methods have a great potential to replace standard methods which are often expensive, time-consuming, and complicated; hence, there is an urgent need to develop an affordable, simple and rapid EC  bioassays/biosensors for analysis of tumor biomarkers. The aim of this doctoral thesis is to develop and optimize bioassays for the detection of such biomarkers, mostly based on nucleic acids, i.e. DNA and RNA. The EC readout will be coupled to isothermal amplification techniques, such as LAMP, RPA or RCA, which rapidly amplify nucleic acids at constant temperatures to improve sensitivity and specificity of detection. Here is the list of selected topics anticipated to be studied in this doctoral thesis: (a) Analysis of DNA mutations in important oncogenes or tumor suppressor genes, implicated in cancer, (b) Analysis of epigenetic modifications, e.g. DNA methylation or upregulated non-coding RNAs, especially microRNAs and long non-coding RNAs, which play a major role in the carcinogenesis process, (c) Application of novel amplification techniques for detection of ultralow levels of nucleic acids, (d) Determination of circulating nucleic acids in body fluids for non-invasive diagnostics, or (e) other similar topics depending on the laboratory needs. These may include but are not limited to development of similar bioassays utilizing electrochemiluminescent (ECL) readout on electrode chips as another promising alternative, or application of third-generation Nanopore sequencing technology. The developed bioassays will be applied to biological and clinical samples and validated with standard methods. The work will be carried out in the Laboratory of Bioelectrochemistry at RECAMO, which is a part of the Masaryk Memorial Cancer Institute.

Supervisor

Mgr. Martin Bartošík, Ph.D.

Proteomická analýza je jedním z klíčových faktorů umožňujících zásadní pokrok našeho poznání nejen v oblasti molekulární biologie a biochemie. Proteomická analýza je souborem jednotlivých kroků zahrnujících přípravu vzorku, jeho vlastní analýzu, převážně s využitím hmotnostní spektrometrie (MS), zpracování naměřených dat, jejich statistické hodnocení a bioinformatickou analýzu, přičemž každý z kroků má zásadní vliv na kvalitu získaných výstupů. Cílem disertační práce v rámci tohoto výzkumného zaměření bude vývoj nových postupů pro proteomickou analýzu historických vzorků, které vyžadují specifický přístup, jak při přípravě vzorků před MS analýzou, tak i při zpracování naměřených MS/MS dat. Studenti budou mít přístup k nejmodernější MS instrumentaci a možnost zapojení do projektů VS Proteomika (CEITEC MU) i spolupracovat s výzkumnými týmy v rámci MU i mimo ni.

Notes

Před podáním přihlášky je nutno se neformálně seznámit s tématem, kontaktujte prof. Zbyňka Zdráhala.

Supervisor

prof. RNDr. Zbyněk Zdráhal, Dr.

Plants employ a broad spectrum of reproductive strategies, ranging from asexual species to hermaphroditism and the presence of distinct sexes. This variety significantly impacts genome architecture. Our objective is to examine plant species with varying reproductive strategies and investigate their responses to e.g. environmental changes, encompassing both biotic and abiotic stresses. Furthermore, we aim to explore the relationship between reproductive modes and genome size, genome dynamics, and ploidy levels. Our research will utilize a wide array of cutting-edge techniques in both forward and reverse genomics, including advanced microscopy and bioinformatics analyses.

Supervisor

RNDr. Roman Hobza, Ph.D.

Navrhovaný výzkum bude reagovat na potřeby současného pokroku v proteomice, glykomice a biomedicíně, který vyžaduje zavedení nových metod, které mohou přinést nové poznatky o proteinech a jejich komplexních systémech. Ve výzkumu chceme využít výhod vlastností elektrochemických přístupů k studiu proteinů a jejich komplexů na nabitých mezifázích. Plán výzkumu vychází ze současných výsledků práce v laboratoři Biofyzikální chemie a molekulární onkologie Biofyzikálního ústavu AV ČR. Budou navrženy a rozvíjeny nové elektrochemické přístupy studia biomedicínsky důležitých proteinů v komplexech s ligandy i peptidy a proteiny s cílem přispět ke stávajícím znalostem o dynamice proteinových komplexů na nabitých mezifázích. Z proteinů, budou zkoumány i glykoproteiny s cílem získání nových informací o proteinové a glykanové části intaktních a chemicky modifikovaných glykoproteinů.

Supervisor

RNDr. Veronika Ostatná, Ph.D.

Recent advances in plant genetic engineering allow precise modifications in desirable genomic region. These methodical approaches are currently employed by both basic researchers and applied biotechnologist to understand complex molecular mechanisms as well as to improve the traits of crop plants. While tools for genetic engineering, such as CRISPR/Cas9, are available for model organisms and most important economically important crops, there are still numerous plant species where precise genetic applications remain complicated or even unfeasible.

This research project will address the identification and overcoming the barriers for successful and high-throughput application of genetic engineering tools in non-model species (including Humulus lupulus, Lotus corniculatus, selected cereal crops etc).

Supervisor

Ing. Vojtěch Hudzieczek, Ph.D.

Our workgroup is interested in molecular basis of severe dyslipidemias in human. The most common of dyslipidemias is familial hypercholesterolemia (FH). The frequency of FH in most populations is about 1/200, and so it is possible to predict that about 50,000 people could be affected in the Czech Republic. The clinical phenotype of FH is caused predominantly by mutations in the LDLR gene. LDLR mutations have been reported along the whole length of the gene. Our workgroup focuses on functional assays of LDLR mutations. For further details please refer to our publications (PMIDs: 27175606, 20663204, 28379029, …).

Supervisor

Mgr. Lukáš Tichý, Ph.D.

Genome sequencing brings a huge amount of information regarding the genetic basis of life. While this information provides a foundation for our understanding of biology, it has become clear that the DNA code alone does not hold all the answers. Epigenetic modifications and higher order DNA structures beyond the double helix contribute to basic biological processes and maintaining cellular stability. Local alternative DNA structures are known to exist in all organisms. Negative supercoiling induces in vitro local nucleotide sequence-dependent DNA structures such as cruciforms, left-handed DNA, triplex and quadruplex structures etc. The formation of cruciforms requires perfect or imperfect inverted repeats of 6 or more nucleotides in the DNA sequence. Inverted repeats are distributed nonrandomly in the vicinity of breakpoint junctions, promoter regions, and at sites of replication initiation. Cruciform structures could for example affect the degree of DNA supercoiling, the positioning of nucleosomes in vivo, and the formation of other secondary structures of DNA. The three-dimensional molecular structure of DNA, specifically the shape of the backbone and grooves of genomic DNA, can be dramatically affected by nucleotide changes, which can cause differences in protein-binding affinity and phenotype. The recognition of cruciform DNA seems to be critical not only for the stability of the genome, but also for numerous, basic biological processes. As such, it is not surprising that many proteins have been shown to exhibit cruciform structure-specific binding properties [1] or G-quadruplex binding properties [2]. Contemporary we have developed easy accessible web tools for analyses of inverted repeats [3] and G-quadruplexes[4] and we have analyzed the presence of inverted repeats and G-quadruplexes in various genomic datasets, such as all sequences mitochondrial genomes [5], all bacterial genomes [6], in S.cerevisiae (in review), in human genome etc. A deeper understanding of the processes related to the formation and function of alternative DNA structures will be an important component to consider in the post-genomic era.

Supervisor

prof. Mgr. Václav Brázda, Ph.D.

Telomeres are the physical ends of linear chromosomes that protect these ends against erroneous recognition as unrepaired chromosomal breaks and regulate the access to Telomerase, a reverse transcriptase that solves the problem terminal DNA loss in each cell cycle. Telomeric structures are known to be composed of short repetitive DNA sequences (telomeric repeats), histone octamers, and number of proteins that bind telomeric DNA, either directly or indirectly, and together, form the protein telomere cap.

Interestingly, telomeric repeats are not exclusively located at the chromosome ends, but they belong among cis-regulatory elements present in promoters of several genes. The distribution of short telomeric motifs (telo-boxes) within the genome is not random, and proteins associated with these telomeric repeats may serve as the epigenetic regulatory mechanisms facilitating metastable changes in gene activity.

The telomeric cap proteins of diverse organisms are less conserved than one might expect. In plants, knowledge of telomere-associated proteins associated with telomeres and regulation of access to telomerase complex is incomplete. The research aims to elucidate the roles of candidate proteins involved in telomerase biogenesis in plants. The outcomes contribute to the characterization of new telomere- or telomerase-associated proteins, complete our knowledge of telomerase assembly or telomere maintenance in plants. In addition, we would like to examine the regulatory factors associated with the telo-boxes present in promoters of the genes active during plant development.

Notes

Poznámky: Práce může být vypracována ve slovenštině či angličtině.

Supervisor

doc. Mgr. Petra Procházková Schrumpfová, Ph.D.

Histone sequence variants and their post-translational modifications (PTMs) are epigenetic marks that significantly influence a number of processes, including the cell cycle and protein interactions. The diversity of histones and the complexity of their modifications in amino acid sequences make histone characterization challenging. The aim of this research is to develop and refine methodologies for the characterization of histone variants and PTMs for mass spectrometry analysis, which will subsequently be used in projects focused on epigenetic regulation in plants, mammals and humans. Epigenetic changes in histones will be investigated in the context of proteome of related cellular signaling pathways.

Supervisor

Mgr. Gabriela Lochmanová, Ph.D.

Our lab is interested in the chromatin structure and dynamics. The chromatin structure must be not only maintained through the cell cycle, but also dynamically modulated during processes like mitosis and replication. Amongst the chromatin-associated complexes, the SMC (Structural Maintenance of Chromosomes) complexes play the central role. Two of them, Cohesin and Condensin, facilitate chromosome segregation and condensation, respectively. Third, the most enigmatic SMC5/6 complex is involved in the DNA damage repair and replication restart, however its essential chromatin-modulating function is still unclear. Our laboratory focuses on the SMC5/6 architecture and functions using state-of-the-art structural biology approaches and various molecular biology tools. For further details please refer to our website (http://www.ncbr.muni.cz/SPEC/) and our publications (https://orcid.org/0000-0002-6223-5169).

Supervisor

doc. Mgr. Jan Paleček, Dr. rer. nat.

Supervisor

doc. RNDr. Miroslav Fojta, CSc.

This research direction includes the structure, evolution and maintenance of telomeres and their roles in chromosome stability, DNA repair and plant speciation. A special attention is given to characterisation of telomerase components and interactors.
Further, we investigate epigenetic mechanisms in the regulation of gene expression, chromatin assembly, genome stability and telomere homeostasis. Biochemical, bioinformatic and molecular biology approaches are applied in this research. As model systems, we primarily use plants and plant cell cultures.
For more details, see our web pages: https://www.ceitec.eu/chromatin-molecular-complexes-jiri-fajkus/rg51

Supervisor

prof. RNDr. Jiří Fajkus, CSc.

Notes

Před podáním přihlášky je vhodné se seznámit s konkrétními tématy pro daný kalendářní rok. Kontakt: doc. Hrstka, MOÚ, Brno.

Supervisor

doc. Mgr. Roman Hrstka, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website , all interested candidates are required to contact Gabriel Demo (gabriel.demo@ceitec.muni.cz) for informal discussion.
Annotation: 
Bunyaviruses are medically important negative-strand RNA viruses whose replication and transcription are mediated by the multifunctional RNA-dependent RNA polymerase (L protein) and viral nucleoproteins (NPs). Recent cryo-electron microscopy studies have revealed the molecular architecture and conformational dynamics of bunyaviral L proteins during RNA synthesis, yet the mechanisms coordinating viral transcription with host translation remain poorly understood. Emerging evidence suggests that viral replication-transcription complexes may localize in proximity to host ribosomes and translation factors, potentially facilitating efficient viral gene expression.
The primary aim of this project is to determine how bunyavirus L proteins, nucleoproteins, and ribonucleoprotein complexes interact with host translational machinery during infection. Initial studies will employ sucrose-gradient polysome profiling, immunoblotting, and mass spectrometry to identify associations between viral replication-transcription factors and host ribosomes or translation factors. Purified viral and host components will be used in biochemical binding assays to validate direct interactions and define minimal interaction networks.
To investigate these interactions under physiologically relevant conditions, a minimal bunyavirus replication-transcription system will be established in mammalian cell lines using viral minigenome approaches. Structural characterization of identified complexes will be pursued using single-particle cryo-electron microscopy and cryo-electron tomography, enabling visualization of viral ribonucleoproteins and associated host translation components. These studies will provide mechanistic insight into how bunyaviruses coordinate RNA synthesis and translation, potentially uncovering novel targets for antiviral intervention.
Recommended literature:

1. Wang X. et al. Structure of Rift Valley Fever Virus RNA-Dependent RNA Polymerase. Journal of Virology (2022).
2. Arragain B. et al. Pre-initiation and elongation structures of full-length La Crosse virus polymerase reveal functionally important conformational changes. Nature Communications (2020).
3. Arragain B. et al. Structural snapshots of La Crosse virus polymerase reveal novel insights into replication and transcription mechanisms. Nature Communications (2022).

 Requirements for candidate:
We are seeking a highly motivated PhD candidate with an MSc degree in structural biology, biochemistry, virology, or a related field.
 
Preferred qualifications:

• Experience in molecular biology, biochemistry, or virology.
• Practical laboratory experience with protein purification, human cell cultures, or RNA biology.
• Basic knowledge of structural biology techniques, particularly cryo-electron microscopy.
• Interest in host-pathogen interactions and viral gene expression.
• Ability to work independently and collaboratively in an interdisciplinary environment.
• Good written and spoken English.

 
PLEASE NOTE: before initiating the formal application process to doctoral studies, all interested candidates are required to contact Gabriel Demo (gabriel.demo@ceitec.muni.cz) for informal discussion.

Supervisor

Mgr. Gabriel Demo, Ph.D.

Annotation: 
Small RNAs are master regulators of gene expression in animals and plants. They guide Argonaute proteins to target RNAs, forming effector complexes that execute RNA silencing - an essential process for cellular homeostasis, development, and defense. Despite decades of research, the molecular principles governing Argonaute activation, guide-RNA loading, and strand selection remain poorly understood.
This PhD project aims to decipher the molecular mechanism of Argonaute loading using state-of-the-art electron cryomicroscopy (cryo-EM) combined with complementary biochemical and functional analyses. Building on our recent discoveries and concepts, we propose that two distinct loading pathways operate in mammals: one orchestrated by Dicer and another by HSP90–co-chaperone systems, both relying on negatively charged intrinsically disordered regions (IDRs) that have been largely overlooked in previous structural studies.
The student will determine high-resolution cryo-EM structures of key intermediates in these pathways-including Dicer-Argonaute-RNA and HSP90-co-chaperone-Argonaute-RNA assemblies-to visualize how conformational changes enable RNA transfer and strand selection.
By resolving this long-standing mechanistic puzzle, the project will define the molecular principles of Argonaute loading, shed light on the evolution and regulation of RNA silencing, and provide structural insights into the pathogenic mechanisms underlying AGO-related developmental disorders (AGO Syndrome).
Requirements for candidate:
Biochemistry/molecular biology/structural biology

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340

Recommended literature:
1) Dicer structure and function: conserved and evolving features. Zapletal D, Kubicek, K, Svoboda P, Stefl R EMBO Reports (2023) 24:e57215 doi:10.15252/embr.202357215
2) microRNAs in action: biogenesis, function and regulation. Shang R, Lee S, Senavirathne G, Lai EC. Nat Rev Genet. 2023 doi:10.1038/s41576-023-00611-y.

Notes

Recommended literature:

1) Dicer structure and function: conserved and evolving features. Zapletal D, Kubicek, K, Svoboda P, Stefl R EMBO Reports (2023) 24:e57215 doi:10.15252/embr.202357215

2) microRNAs in action: biogenesis, function and regulation. Shang R, Lee S, Senavirathne G, Lai EC. Nat Rev Genet. 2023 doi:10.1038/s41576-023-00611-y.

Supervisor

prof. Mgr. Richard Štefl, Ph.D.

Annotation:
To initiate infection, viruses deliver their genomes into host cells. Whereas enveloped viruses fuse their membrane with that of a cell, the cell entry mechanisms employed by non-enveloped viruses are less understood. Recently, it has been shown that endosome rupture enables cell entry of picornaviruses. The student will analyze the putative role of endosome rupture in the cell entry of adenoviruses, polyomaviruses, and parvoviruses. He/She will employ cryo-electron microscopy and tomography to visualize the early stages of cell virus entry in peripheral parts of cells that can be imaged using transmission electron microscopy. The student will analyze changes in the structure of virus particles and endosome membranes that enable the viruses to deliver their genomes into the cytoplasm.
Requirements for candidate:
The prospective student should be interested in learning cryo-EM and structure determination approaches. Previous experience with molecular biology, programming, scripting, and data analyses is a plus.


PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340
Recommended literature:
Virus entry by endocytosis. Mercer J, Schelhaas M, Helenius A. Annu Rev Biochem. 2010;79:803-33. doi: 10.1146/annurev-biochem-060208-104626. PMID: 20196649
 
Adenovirus Entry: From Infection to Immunity. Greber UF, Flatt JW. Annu Rev Virol. 2019 Sep 29;6(1):177-197. doi: 10.1146/annurev-virology-092818-015550. Epub 2019 Jul 5. PMID: 31283442
 
Sending mixed signals: polyomavirus entry and trafficking. Mayberry CL, Bond AC, Wilczek MP, Mehmood K, Maginnis MS. Curr Opin Virol. 2021 Apr;47:95-105. doi: 10.1016/j.coviro.2021.02.004. Epub 2021 Mar 6. PMID: 33690104
 
Parvoviral host range and cell entry mechanisms. Cotmore SF, Tattersall P. Adv Virus Res. 2007;70:183-232. doi: 10.1016/S0065-3527(07)70005-2. PMID: 17765706

Supervisor

doc. Mgr. Pavel Plevka, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website
Annotation: 
Cyclin-dependent kinase 11 (CDK11) is ubiquitously expressed in all tissues indicating an important role for CDK11 in cells. Our recent studies has shown CDK11 plays key roles in transcription of replication-dependent histone genes and in co-transcriptional mRNA splicing (1, 2). Notably, several recent studies identified CDK11 as a candidate essential gene for growth of several cancers therefore, understanding the molecular mechanism(s) of CDK11-dependent gene expression is also of significant clinical interest. In this research we will use CDK11 inhibitors and advanced techniques of molecular biology and biochemistry to characterize roles of CDK11 in regulation of gene expression and tumorigenesis.
Requirements for candidate:
Background in molecular biology, biochemistry or life sciences. Interest in bioinformatics and data analyses is desirable.
Recommended literature:
1) Hluchy M, Gajduskova, P., Ruiz de los Mozos I., Rajecky M., Kluge M., Berger BT., Slaba Z. Potesil D., Weis E., Ule J., Zdrahal Z., Knapp S., Paruch K., Friedel CC., Blazek D*. CDK11 regulates pre-mRNA splicing by phosphorylation of SF3B1. Nature; 609(7928):829-834 (2022)
2) Gajduskova, P., Ruiz de Los Mozos I, Rajecky M., Hluchy M., Ule J., Blazek D*: CDK11 is required for transcription of replication dependent histone genes. Nature Structural & Molecular Biology  27 (5):500-510 (2020).

Supervisor

Mgr. Dalibor Blažek, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website
Annotation:  The development and progression of B-cell malignancies are critically influenced by signals originating from the tumor microenvironment. Over the past decade, non-coding RNAs have emerged as key regulators of cellular communication and signaling pathways in cancer. While microRNAs have been extensively studied and are known to play important roles in B-cell biology, the functions of long non-coding RNAs (lncRNAs) remain largely unexplored. Understanding how lncRNAs regulate malignant B-cell behavior represents one of the major unanswered questions in contemporary cancer biology.
This PhD project aims to uncover the role of lncRNAs in controlling B-cell receptor (BCR) signaling and B–T cell interactions, two fundamental processes that drive the pathogenesis of chronic lymphocytic leukemia (CLL). The project builds on the long-standing expertise of the research group in non-coding RNA biology and tumor–microenvironment interactions, supported by an ERC Starting Grant and multiple high-impact publications in the field. Preliminary data have identified several previously uncharacterized lncRNAs that are likely involved in regulating communication between CLL cells and their microenvironment.
The PhD candidate will investigate the molecular functions of these lncRNAs using a combination of state-of-the-art genetic, cellular, and biochemical approaches. A unique aspect of the project is the availability of newly generated lncRNA knockout mouse models, including mice carrying the genetic deletion of a candidate lncRNA. The student will characterize these models and combine them with established CLL mouse systems, including the Eu-TCL1 model, to determine the role of lncRNAs in leukemia development and progression in vivo. The project will further employ CRISPR interference technologies, RNA pulldown assays, transcriptomic analyses, and studies of primary patient-derived samples to identify molecular pathways and interaction partners controlled by candidate lncRNAs. In parallel, the student will take advantage of a recently developed co-culture system that enables robust proliferation of primary CLL cells in vitro, overcoming a major limitation in CLL research. This innovative platform will be used to perform the first CRISPR-based functional screens aimed at identifying lncRNAs and protein-coding genes that regulate the proliferation and survival of primary CLL cells. By integrating functional genomics, mouse modeling, and patient-based research, this project seeks to establish a comprehensive understanding of how lncRNAs shape B-cell signaling and microenvironmental responses in CLL. The findings are expected to reveal fundamental mechanisms of leukemia biology, identify novel therapeutic vulnerabilities, and generate insights that are broadly relevant to other B-cell malignancies, autoimmune diseases, and normal immune regulation.
This interdisciplinary project offers extensive training in molecular biology, cancer genomics, genome engineering, mouse models, and translational research, providing an excellent foundation for a scientific career in cancer and immunology research.
Requirements on candidates:

• Motivated smart people that have the “drive” to work independently, but also willing to learn from other people in the lab and collaborate.
• Candidates should have a master’s degree in Molecular biology, Biochemistry, or similar field and have deep interest in molecular biology and cancer cell biology.

Supervisor

prof. MUDr. Mgr. Marek Mráz, Ph.D.

Annotation: 
Co-translational quality control is triggered as a response to translational stalling events. Yet, different molecular mechanisms are employed for the recognition of these stalls and to trigger downstream rescue and quality control pathways. The use of collided ribosomes as a proxy for the recognition of translation problems in the cell is conserved from bacteria to humans. In eukaryotes, co-translational quality-control processes triggered by ribosome collisions accomplish several tasks and eventually trigger stress response signalling pathways. These tasks include the degradation of aberrant mRNAs, the degradation of potentially deleterious nascent peptides, the ribosomal subunit rescue and tRNA recycling. We mainly use structural analysis by cryo-EM to gain mechanistic understanding of these translational control events. To that end, we reconstitute macromolecular complexes involved in these processes in vitro or isolate them from cells.
The successful candidate will utilize a multidisciplinary approach to provide detailed mechanistic understanding of the critical human co-translational processes. He/she will utilize human cell cultures, protein expression and purification techniques and biochemistry methods to produce samples for cryogenic electron microscopy (cryo-EM). Comprehensive training in cryo-EM will be available to the successful candidate.

Requirements for candidate:
The ideal candidate should have background in either molecular biology, biochemistry or structural biology. Experience with human cell culture work, RNA biochemistry or protein expression and purification is a strong plus.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340

Recommended literature:
1. Filbeck, S., et al., Ribosome-associated quality-control mechanisms from bacteria to humans. Mol Cell, 2022. 82(8): p. 1451-1466.
2. Ikeuchi, K., et al., Collided ribosomes form a unique structural interface to induce Hel2-driven quality control pathways. EMBO J, 2019. 38(5).
3. Saito, K., et al., Ribosome collisions induce mRNA cleavage and ribosome rescue in bacteria. Nature, 2022. 603(7901): p. 503-508.
4. Narita, M., et al., A distinct mammalian disome collision interface harbors K63-linked polyubiquitination of uS10 to trigger hRQT-mediated subunit dissociation. Nat Commun, 2022. 13(1): p. 6411.
5. Wu, C.C., et al., Ribosome Collisions Trigger General Stress Responses to Regulate Cell Fate. Cell, 2020. 182(2): p. 404-416 e14.
 

Supervisor

RNDr. Petr Těšina, Ph.D.

Annotation:
 
Tick-borne encephalitis virus (TBEV) is a medically significant flavivirus causing severe neurological disease in humans. Despite the availability of TBE vaccines, the number of TBE cases continues to rise, necessitating further research into the immune response to TBEV infection to enable the development of therapeutics. The student will use cryo-electron microscopy to determine the structures of antibody-TBEV complexes at near-atomic resolution, revealing binding modes and conformational changes to both the virus envelope proteins and antibodies. Using EMPEM approach the student will analyse polyclonal antibodies from human sera and compare their epitopes to those of functionally well-characterized  mabs with different neutralizing and/or enhancing properties.
Requirements for candidate:
The prospective student should be interested in learning cryo-EM and structure determination approaches. Previous experience with molecular biology, programming, scripting, and data analyses is a plus.
PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340
Recommended literature:
Torrents de la Peña A, Ward AB. Microfluidics combined with electron microscopy for rapid and high-throughput mapping of antibody-viral glycoprotein complexes. Sewall LM, de Paiva Froes Rocha R, Gibson G, Louie M, Xie Z, Bangaru S, Tran AS, Ozorowski G, Mohanty S, Beutler N, Rogers TF, Burton DR, Shaw AC, Batista FD, Chocarro Ruiz B, Nat Biomed Eng. 2025 Jun 3:10.1038/s41551-025-01411-x. doi: 10.1038/s41551-025-01411-x. Epub ahead of print. PMID: 40461656; PMCID: PMC12404239.
 
Fuzik T, Formanova P, Ruzek D, Yoshii K, Niedrig M, Plevka P. Structure of tick-borne encephalitis virus and its neutralization by a monoclonal antibody. Nat Commun. 2018;9(1):436. doi: 10.1038/s41467-018-02882-0.
 
The structure of immature tick-borne encephalitis virus supports the collapse model of flavivirus maturation.
Anastasina M, Füzik T, Domanska A, Pulkkinen LIA, Šmerdová L, Formanová PP, Straková P, Nováček J, Růžek D, Plevka P, Butcher SJ. Sci Adv. 2024 Jul 5;10(27):eadl1888. doi: 10.1126/sciadv.adl1888.
 
Kuhn RJ, Zhang W, Rossmann MG, Pletnev SV, Corver J, Lenches E, et al. Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell. 2002;108(5):717-25. doi: 10.1016/s0092-8674(02)00660-8.

Supervisor

doc. Mgr. Pavel Plevka, Ph.D.

Annotation: 
 
CDK12 is a transcriptional cyclin-dependent kinase (CDK) found mutated in various cancers. In previous studies, we found that CDK12 maintains genome stability via optimal transcription of key homologous recombination repair pathway genes, including BRCA1, and plays a role in cell cycle progression by regulating processivity of RNA Polymerase IIat core DNA replication genes. Apart from the C-terminal domain of RNA Polymerase II, other cellular substrates of CDK12 are not known. In this research, we propose using a screen in cells carrying an analogsensitive mutant of CDK12 to discover its novel cellular substrates. The substrates and their roles in normal and cancerous cells will be characterized by advanced techniques of molecular biology and biochemistry.

Requirements for candidate:
 
Background in molecular biology, biochemistry, or life sciences. Interest in bioinformatics and data analysis is desirable.
PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340

Recommended literature:
 
1. Pilarova K, Herudek J, Blazek D.*: CDK12: Cellular functions and therapeutic potential of versatile player in cancer: Nucleic Acids Research Cancer (Oxford University Press) k2 (1): zcaa003 (2020)
2. Chirackal Manavalan A.P., Pilarova K., Kluge M., Bartholomeeusen K., Oppelt J., Khirsariya P., Paruch K., Krejci L., Friedel C.C., Blazek D* : CDK12 controls G1/S progression via regulating RNAPII processivity at core DNA replication genes. EMBO reports 20(9):47592 (2019)
3. Ekumi KM, Paculova H, Lenasi T, Pospichalova V, Bösken CA, Rybarikova J, Bryja V, Geyer M, Blazek D*, Barboric M*. Ovarian carcinoma CDK12 mutations misregulate expression of DNA repair genes via deficient formation and function of the Cdk12/CycK complex. Nucleic Acids Research 43(5):2575-89 (2015)
4. Bösken CA, Farnung L, Hintermair C, Merzel Schachter M, Vogel-Bachmayr K, Blazek D, Anand K, Fisher RP, Eick D, Geyer M. The structure and substrate specificity of human Cdk12/Cyclin K. Nature Communications 5 (2014).

Supervisor

Mgr. Dalibor Blažek, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website
Annotation: 
Paramyxoviruses are membrane-enveloped RNA viruses that include major human pathogens such as measles and mumps viruses, as well as highly lethal emerging zoonotic viruses such as Nipah virus, which causes encephalitis with case fatality rates exceeding 50%. Despite their medical importance, there are currently no specific antiviral therapies targeting paramyxovirus infection.
A promising antiviral target is the process of virus assembly and budding, which is orchestrated by the viral matrix protein. Matrix acts as a molecular scaffold at the host-cell plasma membrane, coordinating the recruitment of viral components and driving membrane remodelling to enable virus release. However, current models do not fully explain how matrix specifically recognises the plasma membrane, selectively engages other viral components, and coordinates the timing of budding from the host cell.
This PhD project will investigate the structure and assembly mechanisms of selected paramyxoviruses both in situ and in vitro, using state-of-the-art cryo-electron microscopy approaches. Single-particle cryo-EM analysis will be used to determine the architecture of purified virions and to define the spatial relationships between individual viral components. Focused ion beam milling, cryo-electron tomography and subtomogram averaging will then be applied to visualise viral assembly and budding directly within virus-producing cells.
These in situ studies will be complemented by in vitro reconstitution of viral components on synthetic lipid membranes. These minimal systems will enable high-resolution cryo-EM analysis of matrix-membrane assemblies and dynamic fluorescence-based studies of assembly behaviour. Together, the project will provide new mechanistic insight into how paramyxoviruses assemble and bud from host cells.
Requirements for candidate:
Applicants should have a strong background in one or more of the following areas or related fields: structural biology, molecular biology, biochemistry, virology or biophysics. Experience with Linux-based systems and scripting would be advantageous, but is not essential.
We are equally interested in candidates who are enthusiastic about science, motivated by the project topic, keen to learn new experimental and computational methods, and driven to tackle important and challenging scientific questions. Hard skills are valuable, but curiosity, commitment and scientific ambition will be valued just as highly.

Recommended literature:
Watkinson RE, Lee B. Nipah virus matrix protein: expert hacker of cellular machines. FEBS Lett. 2016;590(15):2494-2511. doi:10.1002/1873-3468.12272

Norris MJ, Husby ML, Kiosses WB, et al. Measles and Nipah virus assembly: Specific lipid binding drives matrix polymerization. Sci Adv. 2022;8(29):eabn1440. doi:10.1126/sciadv.abn1440

Clemente CM, Mobarec JC, Bharat TAM. Applications and prospects of cryo-electron tomography in drug discovery and understanding disease. Curr Opin Struct Biol. 2026;98:103283. doi:10.1016/j.sbi.2026.103283

Supervisor

Mgr. Dominik Hrebík, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website
Annotation:  B-cell malignancies, including chronic lymphocytic leukemia (CLL) and follicular lymphoma (FL), remain incurable diseases despite major advances in targeted therapies. Increasing evidence indicates that malignant B cells acquire profound alterations in chromatin organization, leading to aberrant activation of enhancers and promoters that drive oncogenic gene-expression programs. These changes create a permissive landscape for transcription factors (TFs), key regulators of cell fate, proliferation, and differentiation, to promote tumor growth and survival. Recent studies suggest that TF activity is further shaped by signals from the tumor microenvironment, including post-translational modifications induced by dysregulated signaling pathways. However, the identity of the critical TFs involved and the mechanisms through which they contribute to disease progression, therapeutic resistance, and interactions with the immune microenvironment remain poorly understood. This PhD project aims to uncover the role of novel transcription factors and chromatin-associated regulators in the pathogenesis of CLL, FL, and related B-cell malignancies. Building on our preliminary data, the PhD candidate will investigate TF-driven regulatory networks controlling malignant B-cell survival, proliferation, and communication with surrounding immune cells. Particular emphasis will be placed on transcriptional programs linked to the oncogene MYC and on identifying opportunities for their therapeutic disruption.
The project combines cutting-edge molecular and functional approaches, including CRISPR-based genome engineering, transcriptomic and epigenomic profiling (RNA-seq, ChIP-seq), analysis of primary patient samples, testing of inhibitors, and studies in advanced in vitro and in vivo mouse models, including in vivo CRISPR screening. The candidate will also evaluate emerging therapeutic strategies targeting transcription factors and chromatin regulators, with the goal of identifying new vulnerabilities that can be exploited for treatment. This interdisciplinary project offers training at the interface of cancer biology, genomics, epigenetics, and translational research, providing an opportunity to contribute to the development of next-generation therapies for B-cell malignancies.
 Requirements for candidate:

• Motivated smart people that have the “drive” to work independently, but also willing to learn from other people in the lab and collaborate.
• Candidates should have a master’s degree in Molecular biology, Biochemistry, or similar field and have deep interest in molecular biology and cancer cell biology.

Recommended literature:
Beekman et al. The reference epigenome and regulatory landscape of chronic lymphocytic leukemia. Nature Medicine 2018
https://pubmed.ncbi.nlm.nih.gov/29785028/
 
Sun et al. The immune microenvironment shapes transcriptional and genetic heterogeneity in chronic lymphocytic leukemia. Blood Advances 2022
https://pubmed.ncbi.nlm.nih.gov/35358998/
 
 

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Supervisor

prof. MUDr. Mgr. Marek Mráz, Ph.D.

Annotation:
Despite decades of study, important aspects of phage replication cycles, such as the mechanism of genome delivery, initiation of head assembly, and genome packaging, are poorly understood. We propose to use cryo-electron microscopy and tomography to characterize replication intermediates of phage LE3 infecting Leptospira. The in situ data collection will be enabled by the dimensions of leptospira cells, which are 100 nm thin. Analysis of the infection intermediates will focus on genome delivery, initiation of head assembly, and genome packaging. These processes cannot be studied in vitro because of the challenges of preparing the corresponding complexes in functional form in sufficient amounts.
Requirements for candidate:
The prospective student should be interested in learning cryo-EM and structure determination approaches. Previous experience with molecular biology, programming, scripting, and data analyses is a plus.
PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340
Recommended literature:
Characterization of LE3 and LE4, the only lytic phages known to infect the spirochete Leptospira. Schiettekatte O, Vincent AT, Malosse C, Lechat P, Chamot-Rooke J, Veyrier FJ, Picardeau M, Bourhy P. Sci Rep. 2018 Aug 6;8(1):11781. doi: 10.1038/s41598-018-29983-6.
 
Molecular architecture of tailed double-stranded DNA phages. Fokine A, Rossmann MG. Bacteriophage. 2014 Jan 1;4(1):e28281. doi: 10.4161/bact.28281. Epub 2014 Feb 21. PMID: 24616838
 
A century of the phage: past, present and future. Salmond GP, Fineran PC. Nat Rev Microbiol. 2015 Dec;13(12):777-86. doi: 10.1038/nrmicro3564. Epub 2015 Nov 9. PMID: 26548913
 
Viral genome packaging machines: Structure and enzymology. Catalano CE, Morais MC. Enzymes. 2021;50:369-413. doi: 10.1016/bs.enz.2021.09.006. Epub 2021 Nov 10. PMID: 34861943
 
Casjens, S. R. (2011). The DNA-packaging nanomotor of tailed bacteriophages. Nature Reviews Microbiology, 9(9), 647–657. doi:10.1038/nrmicro2632

Supervisor

doc. Mgr. Pavel Plevka, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website
Annotation: 
The bottom-up construction of an autonomously dividing minimal cell would represent a major advance in synthetic cell biology and provide a stringent test of our understanding of the fundamental principles of life, particularly the mechanisms that drive cell division. Decades of research have established Bacillus subtilis as a model organism for studying cell division in Gram-positive bacteria. However, despite substantial progress, a high-resolution structure of the bacterial division machinery remains elusive.
This PhD project will investigate the structure and assembly mechanism of the bacterial division Z-ring both in situ and in vitro, using a combination of focused ion beam milling, cryo-electron tomography, subtomogram averaging and single-particle cryo-EM analysis. The student will first optimise strategies to arrest cell division at a defined stage, enabling Z-ring structures to be averaged across multiple cells. They will then develop an approach for orienting cells perpendicular to the cryo-EM grid, facilitating downstream FIB-milling and high-resolution cryo-electron tomography.These methods will be used to determine the architecture of the Z-ring in its native cellular context.
In parallel, the student will support the in situ observations with in vitro reconstitutions of the division complex on artificial membranes, followed by structural analysis using cryo-EM. Together, these approaches will provide new insight into the molecular organisation and assembly of the bacterial cell division machinery, with broader implications for the design of antibiotics targeting cell division, synthetic cell biology and the design of minimal dividing cells.
Requirements for candidate:
Applicants should have a strong background in one or more of the following areas or related fields: structural biology, molecular biology, biochemistry or biophysics. Experience with Linux-based systems and scripting would be advantageous, but is not essential.
We are equally interested in candidates who are enthusiastic about science, motivated by the project topic, keen to learn new experimental and computational methods, and driven to tackle important and challenging scientific questions. Hard skills are valuable, but curiosity, commitment and scientific ambition will be valued just as highly.
Recommended literature:
Cameron TA, Margolin W. Insights into the assembly and regulation of the bacterial divisome. Nat Rev Microbiol. 2024;22(1):33-45. doi:10.1038/s41579-023-00942-x

Clemente CM, Mobarec JC, Bharat TAM. Applications and prospects of cryo-electron tomography in drug discovery and understanding disease. Curr Opin Struct Biol. 2026;98:103283. doi:10.1016/j.sbi.2026.103283

Errington J, Wu LJ. Cell Cycle Machinery in Bacillus subtilis. Subcell Biochem. 2017;84:67-101. doi: 10.1007/978-3-319-53047-5_3. PMID: 28500523; PMCID: PMC6126333.

Klumpe S, Plitzko JM. Cryo-focused ion beam milling for cryo-electron tomography: Shaping the future of in situ structural biology. Curr Opin Struct Biol. 2025;94:103138. doi:10.1016/j.sbi.2025.103138

Supervisor

Mgr. Dominik Hrebík, Ph.D.

Annotation:
Tailed phages are a diverse group of viruses abundant in all econiches as revealed by recent metagenomic studies. In the past ten years a substantial effort has been made over an evolutionary sound classification of these viruses. However, due to rapid evolution, the phylogenetic signal, contained in the amino acid sequence of tailed phages, is eroded. As a result, current classification stays fragmented. On the other hand, protein folds remain conserved over greater evolutionary distances. Therefore, phylogenetic inference based on the structure of phage virion proteins may provide a deeper insight into their evolutionary history. Currently, the methods for structure-guided phylogeny are not well developed. The purpose of this project is to optimize the method for maximum likelihood phylogenetic inference on protein structure and use it to reveal new evolutionary connections within tailed phages. The work will involve general bioinformatics methods, including bash-scripting, database search tools, multiple sequence alignment tools and phylogenetic software. The candidate will gain extensive theoretical background and practical skills to work in the field of evolutionary biology.
Requirements for candidate:
The prospective student should be interested in learning techniques of structural phylogeny. Previous experience with molecular biology, programming, scripting, and data analyses is a plus.
PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website (link).
More information:
https://www.ceitec.eu/admission-step-by-step/t11340
Recommended literature:
Weinheimer AR, Ha AD, Aylward FO. Towards a unifying phylogenomic framework for tailed phages. PLoS Genet. 2025 Feb 5;21(2):e1011595. doi: 10.1371/journal.pgen.1011595. PMID: 39908317; PMCID: PMC11835377.
 
Mifsud JCO, Suchard MA, Holmes EC, Lemey P. Recent advances in the inference of deep viral evolutionary history. J Virol. 2025 Sep 23;99(9):e0029225. doi: 10.1128/jvi.00292-25. Epub 2025 Aug 22. PMID: 40844272; PMCID: PMC12456133.
 
Ng WM, Stelfox AJ, Bowden TA. Unraveling virus relationships by structure-based phylogenetic classification. Virus Evol. 2020 Feb 12;6(1):veaa003. doi: 10.1093/ve/veaa003. PMID: 32064119; PMCID: PMC7015158.
 
Moi D, Bernard C, Steinegger M, Nevers Y, Langleib M, Dessimoz C. Structural phylogenetics unravels the evolutionary diversification of communication systems in gram-positive bacteria and their viruses. Nat Struct Mol Biol. 2025 Oct 10. doi: 10.1038/s41594-025-01649-8. Epub ahead of print. PMID: 41073779.

Supervisor

doc. Mgr. Pavel Plevka, Ph.D.

PLEASE NOTE: Before starting the formal application process, applicants must register on the CEITEC PhD School website

Annotation: 
Every organism faces a constant battle against invading genetic elements such as viruses and plasmids. To survive, bacteria and archaea have evolved a remarkable diversity of immune systems that detect and eliminate foreign DNA. Traditionally, these systems have been studied as independent defense mechanisms. However, recent discoveries suggest that some immune systems cooperate, forming more complex defense networks capable of responding to a broader range of threats.

One particularly intriguing example is provided by prokaryotic Argonaute proteins (pAgos), programmable nucleic-acid recognition factors found throughout bacteria and archaea. While some pAgos directly destroy invading DNA, many are genetically associated with proteins belonging to entirely different immune pathways. Why these systems became linked during evolution and how they cooperate at the molecular level remains unknown.
This PhD project will investigate how Argonaute proteins interact with partner immune systems to create novel defense strategies. Using state-of-the-art electron cryomicroscopy (cryo-EM), the student will determine structures of immune complexes captured at different stages of substrate recognition, remodeling, and processing. These structural studies will be complemented by biochemical, biophysical, microbiological, and evolutionary analyses performed in collaboration with leading international laboratories.
A particular focus will be understanding how catalytically inactive Argonautes have evolved into programmable regulators of partner immune proteins and how such cooperation increases the complexity and robustness of microbial immunity. By visualizing molecular machines in action and uncovering their dynamic mechanisms, the student will reveal fundamental principles governing the evolution and organization of immune systems.
The project sits at the interface of structural biology, microbiology, evolutionary biology, and biophysics, providing training in cryo-EM, protein biochemistry, computational structural biology, and molecular mechanism discovery.
Join us to uncover how evolution creates new immune systems by combining existing ones into more powerful defense machines.

Requirements for candidate:
Biochemistry/molecular biology/strucktural biology
Recommended literature:
1) Koopal B. et al. Diverse prokaryotic Argonaute-associated immune systems. Science (2023).
2) Ugarte R. et al. Molecular mechanisms of Argonaute-associated defense systems. Mol Cell (2025).
3) Makarova KS et al. Evolutionary classification of CRISPR-Cas systems. Nat Rev Microbiol (2020).
4) Swarts DC et al. Prokaryotic Argonautes: mechanisms and biological roles. Nat Rev Microbiol.
5) Finocchio G. et al. Cooperation between prokaryotic immune systems.
 

Supervisor

prof. Mgr. Richard Štefl, Ph.D.