A5064891550- CRISPR and Genetic Engineering
- DNA Repair Mechanisms
- Advanced biosensing and bioanalysis techniques
- RNA regulation and disease
- Genomics and Chromatin Dynamics
- Cancer Genomics and Diagnostics
- DNA and Nucleic Acid Chemistry
- Renal cell carcinoma treatment
- RNA and protein synthesis mechanisms
- Chromosomal and Genetic Variations
- Stress Responses and Cortisol
- Cytomegalovirus and herpesvirus research
- Genetic Neurodegenerative Diseases
- Cancer-related Molecular Pathways
- Vibrio bacteria research studies
- RNA Interference and Gene Delivery
- Muscle Physiology and Disorders
- RNA modifications and cancer
- Photosynthetic Processes and Mechanisms
- Resilience and Mental Health
- Suicide and Self-Harm Studies
- Epigenetics and DNA Methylation
- BRCA gene mutations in cancer
- Plant Molecular Biology Research
- Connexins and lens biology
AstraZeneca (Sweden)
2021-2025
MRC Laboratory of Molecular Biology
2016-2023
Medical Research Council
2016-2023
University of Belgrade
2013-2017
Acute treatment with replication-stalling chemotherapeutics causes reversal of replication forks. BRCA proteins protect reversed forks from nucleolytic degradation, and their loss leads to chemosensitivity. Here, we show that fork degradation is no longer detectable in BRCA1-deficient cancer cells exposed multiple cisplatin doses, mimicking a clinical regimen. This effect depends on increased expression chromatin loading PRIMPOL regulated by ATR activity. Electron microscopy single-molecule...
Abstract Genome editing, specifically CRISPR/Cas9 technology, has revolutionized biomedical research and offers potential cures for genetic diseases. Despite rapid progress, low efficiency of targeted DNA integration generation unintended mutations represent major limitations genome editing applications caused by the interplay with double-strand break repair pathways. To address this, we conduct a large-scale compound library screen to identify targets enhancing insertions. Our study reveals...
Article26 November 2018Open Access Transparent process R-loop formation during S phase is restricted by PrimPol-mediated repriming Saša Šviković MRC Laboratory of Molecular Biology, Cambridge, UK Search for more papers this author Alastair Crisp Sue Mei Tan-Wong Sir William Dunn School Pathology, Oxford, Thomas A Guilliam Genome Damage & Stability Centre, Life Sciences, University Sussex, Brighton, Aidan J Doherty Nicholas Proudfoot Guillaume Guilbaud Julian E Sale Corresponding Author...
Article23 July 2020Open Access Timeless couples G-quadruplex detection with processing by DDX11 helicase during DNA replication Leticia K Lerner MRC Laboratory of Molecular Biology, Cambridge, UK Search for more papers this author Sandro Holzer Department Biochemistry, University Mairi L Kilkenny Saša Šviković Pierre Murat Davide Schiavone Cara B Eldridge Alice Bittleston Joseph D Maman Dana Branzei orcid.org/0000-0002-0544-4888 IFOM, Fondazione Italiana per la Ricerca sul Cancro, Institute...
Abstract Prime editing recently emerged as a next-generation approach for precise genome editing. Here we exploit DNA double-strand break (DSB) repair to develop two strategies that install genomic insertions using an Sp Cas9 nuclease-based prime editor (PEn). We first demonstrate PEn coupled regular guide RNA (pegRNA) efficiently promotes short through homology-dependent DSB mechanism. While leads increased levels of by-products, it can rescue pegRNAs perform poorly with nickase-based...
Prime editing is a genome engineering tool that allows installation of small edits with high precision. However, prime efficiency and purity can vary widely across different edits, genomic targets, cell types. nuclease (PEn) utilizes fully active Cas9 instead the nickase employed in conventional editors. PEn capable sites resistant to nickase-based editors but induces more undesired events. In this work, we introduce two strategies enhance precision efficiency. First, apply molecule...
Prime editors (PEs) employ reverse transcriptase (RT) to install genomic edits using a template within the prime editing guide RNA (pegRNA). RT creates 3' flap containing intended edit. However, transcription can continue beyond template, incorporating pegRNA scaffold sequence into flap. These scaffold-derived by-products be installed alongside edit, reducing precision. Here, we develop method that prevents from accessing scaffold, thereby mitigating such by-products. We demonstrate an...
Objectives. Adenosine to inosine RNA editing, serotonin 2C receptor (HTR2C), and stressful life events (SLEs) have all been implicated in suicidal behaviour. We examined the main moderating effects of editing (ADAR, ADARB1) HTR2C genes, childhood trauma (CT), recent SLEs psychiatric disorders as contributors suicide attempt (SA) vulnerability. Methods. Study included 165 attempters 188 non-attempters, diagnosed with one major disorders. CT were assessed using Early Trauma Inventory–Self...
Abstract The primase/polymerase PRIMPOL restarts DNA synthesis when replication is arrested by template impediments. However, we do not have a comprehensive view of how PRIMPOL-dependent repriming integrates with the main pathways damage tolerance, REV1-dependent ‘on-the-fly’ lesion bypass at fork and PCNA ubiquitination-dependent post-replicative gap filling. Guided genome-wide CRISPR/Cas9 screens to survey genetic interactions in non-transformed p53-proficient human cell line, find that...
How noncoding transcription influences chromatin states is still unclear. The
Summary During DNA replication, conflicts with ongoing transcription are frequent and require careful management to avoid genetic instability. R-loops, three stranded nucleic acid structures comprising a DNA:RNA hybrid displaced single DNA, important drivers of damage arising from such conflicts. How R-loops stall replication the mechanisms that restrain their formation during S phase incompletely understood. Here we show in vivo how R-loop drives short purine-rich repeat, (GAA) 10 , become...
ABSTRACT Genome editing tools, especially CRISPR/Cas9-based strategies, have transformed biomedical research and opened opportunities for developing curative treatments genetic diseases. Despite rapid progress, low efficiency of targeted DNA integration generation undesired mutations represent major limitations genome applications. Both issues arise from the interplay between main Double-Strand Break (DSB) repair pathways, Homology-Directed Repair (HDR), Non-Homologous End Joining (NHEJ),...
How noncoding transcription influences chromatin states is still unclear. The Arabidopsis floral repressor gene FLC quantitatively regulated through an antisense-mediated silencing mechanism. antisense transcripts form a cotranscriptional R-loop that dynamically resolved by RNA 3' processing factors (FCA and FY), this linked to silencing. Here, we investigate mechanism show, using single-molecule DNA fiber analysis, FCA FY are required for unimpeded replication fork progression across the...
Regions of the genome with potential to form secondary structure pose a frequent and significant impediment DNA replication must be actively managed in order preserve genetic epigenetic integrity. The fork protection complex (FPC), conserved group replisome-associated proteins including Timeless, Tipin, Claspin, plays an important role maintaining efficient replisome activation, ensuring optimum rates, sister chromatid cohesion checkpoint function. It also helps maintain stability sequences...
Abstract Prime editing recently emerged as a next-generation approach for precise genome editing. Here we exploit DNA double-strand break (DSB) repair to develop two novel strategies that install genomic insertions using an Sp Cas9 nuclease-based prime editor (PEn). We first demonstrate PEn coupled regular guide RNA (pegRNA) efficiently promotes short through homology-dependent DSB mechanism. While lead increased levels of by-products, it rescued pegRNAs performed poorly with nickase-based...