A5010096006- CRISPR and Genetic Engineering
- Animal Genetics and Reproduction
- Pluripotent Stem Cells Research
- Hemoglobinopathies and Related Disorders
- RNA regulation and disease
- Tardigrade Biology and Ecology
- Biocrusts and Microbial Ecology
- Advanced biosensing and bioanalysis techniques
- Muscle Physiology and Disorders
- Virus-based gene therapy research
- RNA and protein synthesis mechanisms
- RNA Interference and Gene Delivery
- Molecular Biology Techniques and Applications
- Hedgehog Signaling Pathway Studies
- Genomics and Chromatin Dynamics
- Ubiquitin and proteasome pathways
- Developmental Biology and Gene Regulation
- Metabolomics and Mass Spectrometry Studies
- RNA Research and Splicing
- Retinal Development and Disorders
- DNA Repair Mechanisms
- Genetics and Neurodevelopmental Disorders
- Genetics, Aging, and Longevity in Model Organisms
- Xenotransplantation and immune response
- Epigenetics and DNA Methylation
Inserm
2016-2025
Centre National de la Recherche Scientifique
2016-2025
Structure et Instabilité des Génomes
2015-2025
Muséum national d'Histoire naturelle
2014-2024
Sorbonne Université
2017-2023
Centre National pour la Recherche Scientifique et Technique (CNRST)
2022
Institut Cochin
1998-2016
Université Paris Cité
1994-2016
Institut National de Recherche en Santé Publique
2014
Délégation Paris 5
2003-2013
CRISPOR.org is a web tool for genome editing experiments with the CRISPR–Cas9 system. It finds guide RNAs in an input sequence and ranks them according to different scores that evaluate potential off-targets of interest predict on-target activity. The list genomes continuously expanded, more 150 added last two years. CRISPOR tries provide comprehensive solution from selection, cloning expression RNA as well providing primers needed testing activity off-targets. Recent developments include...
Hypoxia-inducible factor 1 (HIF-1) is a basic helix-loop-helix transcription which expressed when mammalian cells are subjected to hypoxia and activates of genes encoding erythropoietin, vascular endothelial growth factor, other proteins that important for maintaining oxygen homeostasis. Previous studies have provided indirect evidence HIF-1 also regulates glycolytic enzymes. In this paper we characterize response elements in the promoters ALDA, ENO1, Ldha genes. We demonstrate plays an...
The success of the CRISPR/Cas9 genome editing technique depends on choice guide RNA sequence, which is facilitated by various websites. Despite importance and popularity these algorithms, it unclear to extent their predictions are in agreement with actual measurements.
Using in vitro amplification of cDNA by the polymerase chain reaction, we have detected spliced transcripts various tissue-specific genes (genes for anti-Müllerian hormone, beta-globin, aldolase A, and factor VIIIc) human nonspecific cells, such as fibroblasts, hepatoma lymphoblasts. In rats, erythroid- liver-type pyruvate kinase were also brain, lung, muscle. The abundance these "illegitimate" is very low; yet, their existence possibility amplifying them reaction provide a powerful tool to...
Sequence-specific nucleases like TALENs and the CRISPR/Cas9 system have greatly expanded genome editing possibilities in model organisms such as zebrafish. Both systems recently been used to create knock-out alleles with great efficiency, also successfully employed knock-in of DNA cassettes at defined loci via homologous recombination (HR). Here we report CRISPR/Cas9-mediated into zebrafish a very high rate by homology-independent double-strand break (DSB) repair pathways. After co-injection...
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....
Abstract Throughout vertebrates, cerebrospinal fluid-contacting neurons (CSF-cNs) are ciliated cells surrounding the central canal in ventral spinal cord. Their contribution to modulate locomotion remains undetermined. Recently, we have shown CSF-cNs by directly projecting onto locomotor pattern generators (CPGs), but sensory modality these convey circuits and their relevance innate remain elusive. Here, demonstrate vivo that form an intraspinal mechanosensory organ detects bending. By...
ABSTRACT Signalling by members of the Hedgehog family secreted proteins plays a central role in development vertebrate and invertebrate embryos. In Drosophila, transduction signal is intimately associated with activity protein kinase A product segment polarity gene patched. We have cloned homologue patched from zebrafish Danio rerio analysed spatiotemporal regulation its transcription during embryonic both wild-type mutant animals. find striking correlation between accumulation patched1...
Gli3 is a zinc finger transcription factor proteolytically processed into truncated repressor lacking C-terminal activation domains. processing stimulated by protein kinase A (PKA) and inhibited Hedgehog signaling, major signaling pathway in vertebrate development disease. We show here that multisite glycogen synthase 3beta (GSK3beta) phosphorylation ubiquitination SCFbetaTrCP are required for processing. identified multiple betaTrCP-binding sites related to the DSGX2-4S motif Gli3, which...
Myogenin, one of the MyoD family proteins, is expressed early during somitogenesis and required for myoblast fusion in vivo . Previous studies transgenic mice have shown that a 184-bp myogenin promoter fragment sufficient to correctly drive expression β-galactosidase transgene embryogenesis. We show here mutation DNA motifs present this region, MEF3 motif, abolished correct transgene. found proteins bind site are homeoproteins Six/ sine oculis family. Antibodies directed specifically against...
Serum response factor (SRF) is at the confluence of multiple signaling pathways controlling transcription immediate-early genes and muscle-specific genes. There are active SRF target sequences in more than 50 expressed three muscle lineages including normal diseased hearts. However, role heart formation has not been addressed vivo thus far due to early requirement for mesoderm formation. We have generated a conditional mutant by using Cre-LoxP strategy that will be extremely useful study...
Muscle fibers show great differences in their contractile and metabolic properties. This diversity enables skeletal muscles to fulfill adapt different tasks. In this report, we that the Six/Eya pathway is implicated establishment maintenance of fast-twitch muscle phenotype. We demonstrate MEF3/Six DNA binding element present aldolase A pM promoter mediates high level activation glycolytic (but not slow-twitch) fibers. also among Six Eya gene products expressed mouse muscle, Six1 Eya1...
A few animal models of Duchenne muscular dystrophy (DMD) are available, large ones such as pigs or dogs being expensive and difficult to handle. Mdx (X-linked dystrophy) mice only partially mimic the human disease, with limited chronic lesions muscle weakness. Their small size also imposes limitations on analyses. rat model could represent a useful alternative since rats animals but 10 times bigger than better reflect functional abnormalities observed in DMD patients. Two lines Dmd...
Abstract Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species. Here we demonstrate that the CRISPR/Cas9 system is highly efficient for editing non-model crop pest Lepidoptera, noctuid moth Spodoptera littoralis . We knocked-out olfactory receptor co-receptor Orco gene to investigate its function olfaction. find 89.6% injected individuals carried mutations, 70% which transmitted them next generation. CRISPR/Cas9-mediated...
Teleost fishes, thanks to their rapid evolution of sex determination mechanisms, provide remarkable opportunities study the formation chromosomes and mechanisms driving birth new master determining (MSD) genes. However, evolutionary interplay between MSD genes they harbor is rather unexplored. We characterized a male-specific duplicate anti-Müllerian hormone (amh) as gene in Northern Pike (Esox lucius), using genomic expression evidence well by loss-of-function gain-of-function experiments....
Chromosomal translocations are signatures of numerous cancers and lead to expression fusion genes that act as oncogenes. The wealth genomic aberrations found in cancer, however, makes it challenging assign a specific phenotypic change aberration. In this study, we set out use genome editing with zinc finger (ZFN) transcription activator-like effector (TALEN) nucleases engineer, de novo, translocation-associated oncogenes at cognate endogenous loci human cells. Using ZFNs TALENs designed cut...
Editing the fetal γ-globin promoters in hematopoietic stem cells from sickle cell disease patients induces therapeutic levels.
Abstract The heartbeat and blood flow signal to endocardial cell progenitors through mechanosensitive proteins that modulate the genetic program controlling heart valve morphogenesis. To date, mechanism by which mechanical forces coordinate tissue morphogenesis is poorly understood. Here we use high-resolution imaging uncover coordinated behaviours leading formation. We find valves originate from located in ventricle atrium generate leaflets a set of movements. Gene profiling analyses live...
Mitochondrial diseases are frequently associated with mutations in mitochondrial DNA (mtDNA). In most cases, mutant and wild-type mtDNAs coexist, resulting heteroplasmy. The selective elimination of mtDNA, consequent enrichment can rescue pathological phenotypes heteroplasmic cells. Use the mitochondrially targeted zinc finger-nuclease (mtZFN) results degradation mtDNA through site-specific cleavage. Here, we describe a substantial enhancement our previous mtZFN-based approaches to targeting...
The generation of gene-edited animals using the CRISPRs/Cas9 system is based on microinjection into zygotes which inefficient, time consuming and demands high technical skills. We report optimization an electroporation method for intact rat sgRNAs Cas9 protein in combination or not with ssODNs (~100 nt). This resulted frequency knockouts, between 15 50% analyzed animals. Importantly, as donor template precise knock-in mutations 25-100% animals, comparable to microinjection. Electroporation...