A5043849446- CRISPR and Genetic Engineering
- DNA Repair Mechanisms
- Genetics, Bioinformatics, and Biomedical Research
- BRCA gene mutations in cancer
- Bioinformatics and Genomic Networks
- Cancer Research and Treatments
- Machine Learning in Bioinformatics
- Genomics and Rare Diseases
- RNA and protein synthesis mechanisms
- interferon and immune responses
- Computational Drug Discovery Methods
- Advanced biosensing and bioanalysis techniques
- Protein Structure and Dynamics
- Health, Environment, Cognitive Aging
- PARP inhibition in cancer therapy
- Nutrition, Genetics, and Disease
- Clay minerals and soil interactions
- thermodynamics and calorimetric analyses
- Epigenetics and DNA Methylation
- Porphyrin Metabolism and Disorders
- Therapeutic Uses of Natural Elements
- Chromosomal and Genetic Variations
- Pluripotent Stem Cells Research
- Genomics, phytochemicals, and oxidative stress
- Genomics and Chromatin Dynamics
Centre National de la Recherche Scientifique
2016-2025
Institut Cochin
2019-2025
Inserm
2019-2025
Université Paris Cité
2019-2025
Institut Gustave Roussy
2015-2023
Université Paris-Saclay
2016-2022
ORCID
2022
Stabilité génétique et oncogenèse
2015-2020
La Ligue Contre le Cancer
2018-2019
Université Paris-Sud
2016-2018
In genome editing with CRISPR-Cas9, transgene integration often remains challenging. Here, we present an approach for increasing the efficiency of by homology-dependent repair (HDR). CtIP, a key protein in early steps homologous recombination, is fused to Cas9 and stimulates HDR at human AAVS1 safe harbor locus. A minimal N-terminal fragment designated HE enhancer, sufficient stimulate this depends on CDK phosphorylation sites multimerization domain essential CtIP activity recombination....
The canonical DNA damage response (DDR) maintains genome stability, involving synthesis/cell cycle arrest. However, unchallenged cells proliferate when they are continually exposed to low-level/endogenous replication stress. We previously discovered and characterized a noncanonical cellular that is specific nonblocking stress, i.e., low-level stress (LoL-DDR), in primary cells. Although this generates stress-induced reactive oxygen species (RIR), it triggers program prevents the accumulation...
Sepiolite nanofibers, which are natural silicates belonging to the clay mineral family, could be promising potential nanocarriers for nonviral transfer of biomolecules. The physicochemical characteristics sepiolite make it capable binding various types biological molecules, including polysaccharides, lipids, proteins and viruses. nanofibers have also been shown bind effectively DNA molecules through electrostatic interactions, hydrogen bonds, cationic bridges van der Waals forces. In this...
<title>Abstract</title> The canonical DNA damage response (cDDR) maintains genome stability, involving synthesis arrest. However, unchallenged cells proliferate when they are continually exposed to low-level/endogenous replication stress. We previously characterized a noncanonical specific nonblocking stress, i.e. low-level stress (LoL-DDR), in primary cells. Although LoL-DDR generates stress-induced ROS (RIR), it prevents the accumulation of premutagenic 8-oxo-guanine (8-oxoG). Primary...
Abstract Sepiolite is a nanofibrous natural silicate that can be used as nanocarrier because it naturally internalized into mammalian cells, due to its nano-size dimension. Therefore, deciphering the mechanisms of sepiolite cell internalization constitutes question interesting biotechnology, for use nanocarrier, well environmental and public health concerns. Though low, perfectly stable intrinsic fluorescence nanofibers allows follow their fate cells by specifically sensitive technics. By...
Replications forks are routinely hindered by different endogenous stresses. Because homologous recombination plays a pivotal role in the reactivation of arrested replication forks, defects reveal initial stress(es). Homologous recombination-defective cells consistently exhibit spontaneously reduced speed, leading to mitotic extra centrosomes. Here, we identify oxidative stress as major source speed deceleration cells. The treatment with antioxidant N-acetyl-cysteine or maintenance at low O2...
Background Primary ovarian insufficiency (POI) affects 1% of women under 40 years and is a public health problem. The genetic causes POI are highly heterogeneous with isolated or syndromic forms. Recently, variants in genes involved DNA repair have been shown to cause POI. Notably, Fanconi anaemia (FA) traits related biallelic BRCA2 truncated has reported. Here, we report novel phenotype variant consanguineous Turkish family. Methods Exome sequencing (ES) was performed the patient. We also...
Abstract Cells are inevitably challenged by low-level/endogenous stresses that do not arrest DNA replication. Here, in human primary cells, we discovered and characterized a noncanonical cellular response is specific to nonblocking replication stress. Although this generates reactive oxygen species (ROS), it induces program prevents the accumulation of premutagenic 8-oxoguanine an adaptive way. Indeed, stress-induced ROS (RIR) activate FOXO1-controlled detoxification genes such as SEPP1,...
Abstract Selection of the appropriate DNA double-strand break (DSB) repair pathway is decisive for genetic stability. It proposed to act according two steps: 1-canonical nonhomologous end-joining (C-NHEJ) versus resection that generates single-stranded (ssDNA) stretches; 2-on ssDNA, gene conversion (GC) nonconservative single-strand annealing (SSA) or alternative (A-EJ). Here, we addressed mechanisms by which RAD51 regulates this second step, preventing in human cells. Silencing BRCA2...
DNA double-strand breaks (DSB) are very harmful lesions that can generate genome rearrangements. In this study, we used intrachromosomal reporters to compare both the efficiency and accuracy of end-joining occurring with close (34 bp apart) vs. distant DSBs (3200 in human fibroblasts. We showed a few kb between two I-SceI-induced sufficient foster deletions capture/insertions at junction scar. Captured sequences mostly coupled be partial duplications reporter (i.e., adjacent DSB) or...
Homologous recombination (HR) is a prominent DNA repair pathway maintaining genome integrity. Mutations in many HR genes lead to cancer predisposition. Paradoxically, the implication of pivotal factor RAD51 on development remains puzzling. Particularly, no mouse models are available address role aging and carcinogenesis vivo. We engineered model with an inducible dominant-negative form (SMRad51) that suppresses RAD51-mediated without stimulating alternative mutagenic pathways. found vivo...
Abstract BRCA1 mutations have been identified that increase the risk of developing hereditary breast and ovarian cancers. Genetic screening is now offered to patients with a family history cancer, adapt their treatment management relatives. However, large number variants uncertain significance (VUS) are detected. To better understand these variants, high-throughput structural functional analysis was performed on set VUS. Information both cellular localization homology-directed DNA repair...
Abstract Endogenous stress represents a major source of genome instability, but is in essence difficult to apprehend. Incorporation labeled radionuclides into DNA constitutes tractable model analyze cellular responses endogenous attacks. Here we show that incorporation [ 3 H]thymidine CHO cells generates oxidative-induced mutagenesis, but, with peak at low doses. Proteomic analysis showed the response differs between and high levels stress. In particular, these results confirmed involvement...
ABSTRACT Primary Ovarian insufficiency (POI) affects 1% of women under forty. We studied a patient with non-syndromic POI, from consanguineous Turkish family. Exome sequencing identified homozygous missense variant c.8524C>T/p.R2842C in BRCA2 . is major player homologous recombination (HR). deficiency induces cancer predisposition and Fanconi Anemia (FA). Remarkably, neither the nor her family exhibit somatic pathologies. The patient’s cells presented intermediate levels chromosomal...
Sepiolite is a natural clay silicate that widely used, including biomedical applications; notably sepiolite shows promising features for the transfer of biological macromolecules into mammalian cells. However, before its use, such an approach should address efficiency binding to and cell toxicity. Because spontaneously forms aggregates, disaggregation can represent important challenge improving suspension performance assembly with species. this also influence toxicity in Here, very pure...
Understanding the effect of an ever-growing number human variants detected by genome sequencing is a medical challenge. The yeast Saccharomyces cerevisiae model has held attention for its capacity to monitor functional impact missense mutations found in genes, including BRCA1 breast and ovarian cancer susceptibility gene. When expressed yeast, wild-type full-length protein forms single nuclear aggregate induces growth inhibition. Both events are modified pathogenic BRCA1. However, biological...
Abstract The DNA damage response (DDR) interrupts cell cycle progression to restore genome integrity. However, unchallenged proliferating cells are continually exposed endogenous stress, raising the question of a stress-threshold for DDR activation. Here, we identified stress threshold below which primary human fibroblasts, activate cell-autonomous that not activates full and arrests progression,. We characterized this “pre-DDR” showing it triggers production reactive oxygen species (ROS) by...
<div>Abstract<p><i>BRCA1</i> mutations have been identified that increase the risk of developing hereditary breast and ovarian cancers. Genetic screening is now offered to patients with a family history cancer, adapt their treatment management relatives. However, large number <i>BRCA1</i> variants uncertain significance (VUS) are detected. To better understand these variants, high-throughput structural functional analysis was performed on set VUS....
<p>Size Exclusion Chromatography profiles obtained on BRCA1 BRCT domains free (red curve) or in complex with BACH1-P (purple), AB-1P (green), AB-2P (blue). The experiment was performed using a Superdex-75 10/300 GL column (GE Healthcare) pre-equilibrated 50mM Tris-HCl pH 7.5, 150 mM NaCl, 10 β-mercaptoethanol and protease inhibitors (Roche).</p>
<p>FBTSA assays performed to measure binding of the mutated BRCA1 BRCT domains ACC1-P, BACH1-P, CtiP-P, AB-1P and AB-2P.</p>
<p>Mutated BRCT domains fused to GST were expressed in E. coli and purified by affinity chromatography using glutathione beads. This figure shows a SDS-PAGE gel with samples from the bacterial pellet (P), supernatant (S) beads (G) after incubation washing. VUS classified into 3 groups as function of amount (1) soluble (2) protein obtained cultures. Lanes corresponding typical insoluble fusion protein, poorly WT are boxed orange, grey black, respectively (colors Figure 4 excepted for...