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CRISPR Journal
Journals -
Establishing the allosteric mechanism in CRISPR‐Cas9
PublicationAllostery is a fundamental property of proteins, which regulates biochemical information transfer between spatially distant sites. Here, we report on the critical role of molecular dynamics (MD) simulations in discovering the mechanism of allosteric communication within CRISPR‐Cas9, a leading genome editing machinery with enormous promises for medicine and biotechnology. MD revealed how allostery intervenes during at least three...
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Principles of target DNA cleavage and the role of Mg2+ in the catalysis of CRISPR–Cas9
PublicationAt the core of the CRISPR–Cas9 genome-editing technology, the endonuclease Cas9 introduces site-specific breaks in DNA. However, precise mechanistic information to ameliorate Cas9 function is still missing. Here, multimicrosecond molecular dynamics, free energy and multiscale simulations are combined with solution NMR and DNA cleavage experiments to resolve the catalytic mechanism of target DNA cleavage. We show that the conformation...
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A CRISPR/Cas9-Based Mutagenesis Protocol for Brachypodium distachyon and Its Allopolyploid Relative, Brachypodium hybridum
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Catalytic Mechanism of Non-Target DNA Cleavage in CRISPR-Cas9 Revealed by Ab Initio Molecular Dynamics
PublicationCRISPR-Cas9 is a cutting-edge genome editing technology, which uses the endonuclease Cas9 to introduce mutations at desired sites of the genome. This revolutionary tool is promising to treat a myriad of human genetic diseases. Nevertheless, the molecular basis of DNA cleavage, which is a fundamental step for genome editing, has not been established. Here, quantum–classical molecular dynamics (MD) and free energy methods are used...
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Discrimination of hospital isolates of Acinetobacter baumannii using repeated sequences and whole genome alignment differential analysis
PublicationAn optimized method for bacterial strain differentiation, based on combination of Repeated Sequences and Whole Genome Alignment Differential Analysis (RS&WGADA), is presented in this report. In this analysis, 51 Acinetobacter baumannii multidrug-resistance strains from one hospital environment and patients from 14 hospital wards were classified on the basis of polymorphisms of repeated sequences located in CRISPR region, variation...
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Novel Tools for Comprehensive Functional Analysis of LDLR (Low-Density Lipoprotein Receptor) Variants
PublicationFamilial hypercholesterolemia (FH) is an autosomal-dominant disorder caused mainly by substitutions in the low-density lipoprotein receptor (LDLR) gene, leading to an increased risk of premature cardiovascular diseases. Tremendous advances in sequencing techniques have resulted in the discovery of more than 3000 variants of the LDLR gene, but not all of them are clinically relevant. Therefore, functional studies of selected variants...
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Molecular Dynamics to Predict Cryo-EM: Capturing Transitions and Short-Lived Conformational States of Biomolecules
PublicationSingle-particle cryogenic electron microscopy (cryo-EM) has revolutionized the field of the structural biology, providing an access to the atomic resolution structures of large biomolecular complexes in their near-native environment. Today’s cryo-EM maps can frequently reach the atomic-level resolution, while often containing a range of resolutions, with conformationally variable regions obtained at 6 Å or worse. Low resolution...
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Genetyka molekularna - wykład, WCh 2022/23, Biotechnologia II stopień - new
e-Learning CoursesCelem kursu jest zapoznanie się z podstawowymi mechanizmami funkcjonowania genomów, regulacji ich aktywności i dziedziczenia, edycją genomu w oparciu o programowane nukleazy (system CRISPR-Cas i prime-editing), metodami wykorzystywanymi do analizy struktury, funkcji genów, skutków zmian mutacyjnych oraz wyłączania ich aktywności (np. TargeTron gene nockout system) oraz technikami in vitro wykorzystywanymi do wprowadzania biomolekuł...
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Genetyka molekularna - wykład, WCh 2023/24, Biotechnologia II stopień
e-Learning CoursesCelem kursu jest zapoznanie się z podstawowymi mechanizmami funkcjonowania genomów, regulacji ich aktywności i dziedziczenia, edycją genomu w oparciu o programowane nukleazy (system CRISPR-Cas i prime-editing), metodami wykorzystywanymi do analizy struktury, funkcji genów, skutków zmian mutacyjnych oraz wyłączania ich aktywności (np. TargeTron gene nockout system) oraz technikami in vitro wykorzystywanymi do wprowadzania biomolekuł...
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Genetyka molekularna - wykład, WCh 2024/25, Biotechnologia II stopień
e-Learning CoursesCelem kursu jest zapoznanie się z podstawowymi mechanizmami funkcjonowania genomów, regulacji ich aktywności i dziedziczenia, edycją genomu w oparciu o programowane nukleazy (system CRISPR-Cas i prime-editing), metodami wykorzystywanymi do analizy struktury, funkcji genów, skutków zmian mutacyjnych oraz wyłączania ich aktywności (np. TargeTron gene nockout system) oraz technikami in vitro wykorzystywanymi do wprowadzania biomolekuł...