A5001141726- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- DNA and Nucleic Acid Chemistry
- Advanced biosensing and bioanalysis techniques
- DNA Repair Mechanisms
- Bacteriophages and microbial interactions
- RNA regulation and disease
- RNA modifications and cancer
- Cytomegalovirus and herpesvirus research
- Enzyme Structure and Function
- RNA Interference and Gene Delivery
- Insect symbiosis and bacterial influences
- Biochemical and Molecular Research
- Plant Virus Research Studies
- Epigenetics and DNA Methylation
- Viral Infections and Immunology Research
- Diffusion and Search Dynamics
- Evolution and Genetic Dynamics
- Force Microscopy Techniques and Applications
- RNA Research and Splicing
- Innovation and Socioeconomic Development
- Mosquito-borne diseases and control
- Lipid Membrane Structure and Behavior
- Ubiquitin and proteasome pathways
Vilnius University
2016-2025
Institute of Biochemistry and Biophysics, Polish Academy of Sciences
2020
Czech Academy of Sciences, Institute of Biotechnology
1999-2015
University of Bristol
2008-2010
University of Liverpool
2010
KU Leuven
2009
University of Edinburgh
2009
Cardiff University
2008
Max Planck Institute of Molecular Cell Biology and Genetics
2005-2008
International Institute of Molecular and Cell Biology
2005-2008
Clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide adaptive immunity against viruses and plasmids in bacteria archaea. The silencing of invading nucleic acids is executed by ribonucleoprotein complexes preloaded with small, interfering CRISPR RNAs (crRNAs) that act as guides for targeting degradation foreign acid. Here, we demonstrate the Cas9–crRNA complex Streptococcus thermophilus CRISPR3/Cas system introduces vitro a double-strand...
The CRISPR/Cas adaptive immune system provides resistance against phages and plasmids in Archaea Bacteria.CRISPR loci integrate short DNA sequences from invading genetic elements that provide small RNA-mediated interference subsequent exposure to matching nucleic acids.In Streptococcus thermophilus, it was previously shown the CRISPR1/Cas can immunity by integrating novel spacers following these foreign subsequently direct specific cleavage of invasive homologous sequences.Here, we show S....
Clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems protect bacteria and archaea from infection by viruses plasmids. Central to this defense is a ribonucleoprotein complex that produces RNA-guided cleavage of foreign nucleic acids. In DNA-targeting CRISPR-Cas systems, the RNA component encodes target recognition forming site-specific hybrid (R-loop) with its complement (protospacer) on an invading DNA while displacing noncomplementary strand....
Type III CRISPR-Cas systems in prokaryotes provide immunity against invading nucleic acids through the coordinated degradation of transcriptionally active DNA and its transcripts by Csm effector complex. The Cas10 subunit complex contains an HD nuclease domain that is responsible for two Palm domains with elusive functions. In addition, Csm6, a ribonuclease not part complex, also required to full immunity. We show here target RNA binding Streptococcus thermophilus triggers synthesize cyclic...
In recent years, CRISPR-associated (Cas) nucleases have revolutionized the genome editing field. Being guided by an RNA to cleave double-stranded (ds) DNA targets near a short sequence termed protospacer adjacent motif (PAM), Cas9 and Cas12 offer unprecedented flexibility, however, more compact versions would simplify delivery extend application. Here, we present collection of 10 exceptionally (422-603 amino acids) CRISPR-Cas12f that recognize dsDNA in PAM dependent manner. Categorized as...
Abstract Transposition has a key role in reshaping genomes of all living organisms 1 . Insertion sequences IS200/IS605 and IS607 families 2 are among the simplest mobile genetic elements contain only genes that required for their transposition its regulation. These encode tnpA transposase, which is essential mobilization, often carry an accessory tnpB gene, dispensable transposition. Although TnpA transposon mobilization well documented, function TnpB remained largely unknown. It had been...
RNA-guided nucleases (RGNs) based on the type II CRISPR-Cas9 system of Streptococcus pyogenes (Sp) have been widely used for genome editing in experimental models. However, nontrivial level off-target activity reported several human cells may hamper clinical translation. RGN specificity depends both guide RNA (gRNA) and protospacer adjacent motif (PAM) recognized by Cas9 protein. We hypothesized that more stringent PAM requirements reduce occurrence mutagenesis. To test this postulation, we...
The Cas9-crRNA complex of the Streptococcus thermophilus DGCC7710 CRISPR3-Cas system functions as an RNA-guided endonuclease with crRNA-directed target sequence recognition and protein-mediated DNA cleavage. We show here that additional RNA molecule, tracrRNA (trans-activating CRISPR RNA), co-purifies Cas9 protein isolated from heterologous E. coli strain carrying S. system. provide experimental evidence is required for Cas9-mediated interference both in vitro vivo. specifically promotes...
To expand the repertoire of Cas9s available for genome targeting, we present a new in vitro method simultaneous examination guide RNA and protospacer adjacent motif (PAM) requirements. The relies on cleavage plasmid libraries containing randomized PAM as function Cas9-guide complex concentration. Using this method, accurately reproduce canonical preferences Streptococcus pyogenes, thermophilus CRISPR3 (Sth3), CRISPR1 (Sth1). Additionally, sgRNA solutions novel Cas9 protein from Brevibacillus...
Abstract Bacterial Cas9 nucleases from type II CRISPR-Cas antiviral defence systems have been repurposed as genome editing tools. Although these proteins are found in many microbes, only a handful of variants used for applications. Here, we use bioinformatic and biochemical analyses to explore this largely uncharacterized diversity. We apply cell-free screens assess the protospacer adjacent motif (PAM) guide RNA (gRNA) requirements 79 proteins, thus identifying at least 7 distinct gRNA...
SpCas9 and AsCas12a are widely utilized as genome-editing tools in human cells. However, their relatively large size poses a limitation for delivery by cargo-size-limited adeno-associated virus (AAV) vectors. The type V-F Cas12f from Acidibacillus sulfuroxidans is exceptionally compact (422 amino acids) has been harnessed tool. Here, we developed an approach, combining deep mutational scanning structure-informed design, to successfully generate two AsCas12f activity-enhanced (enAsCas12f)...
CRISPR-Cas systems provide bacteria and archaea with adaptive immunity against foreign nucleic acids. In type I systems, invading DNA is detected by a large ribonucleoprotein surveillance complex called Cascade. The crRNA component of Cascade used to recognize target sites in (protospacers) formation an R-loop driven base-pairing complementarity. Using single-molecule supercoiling experiments near base-pair resolution, we probe here the mechanism detect short-lived intermediates on...