A5014396815- Muscle Physiology and Disorders
- RNA Research and Splicing
- Tissue Engineering and Regenerative Medicine
- Pluripotent Stem Cells Research
- Mesenchymal stem cell research
- CRISPR and Genetic Engineering
- Ion channel regulation and function
- Pelvic floor disorders treatments
- Genetic Neurodegenerative Diseases
- Cardiomyopathy and Myosin Studies
- RNA modifications and cancer
- CAR-T cell therapy research
- Ubiquitin and proteasome pathways
- Virus-based gene therapy research
- Nicotinic Acetylcholine Receptors Study
- Histone Deacetylase Inhibitors Research
- Growth Hormone and Insulin-like Growth Factors
- 3D Printing in Biomedical Research
- Viral Infections and Immunology Research
- Urological Disorders and Treatments
- Genetic Syndromes and Imprinting
- Neurogenetic and Muscular Disorders Research
- Telomeres, Telomerase, and Senescence
- Animal Genetics and Reproduction
- Molecular Biology Techniques and Applications
Institut des Cellules Souches pour le Traitement et l'Étude des Maladies Monogéniques
2015-2024
Inserm
1993-2024
Association Francaise contre les Myopathies
2024
Centre Hospitalier Sud Francilien
2021-2022
Centre National de la Recherche Scientifique
1988-2021
Genethon (France)
2018
Université d'Évry Val-d'Essonne
2018
Assistance Publique – Hôpitaux de Paris
2013
Hôpital Tenon
2013
Sorbonne Université
2013
Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells living organisms. However, their delivery into target can be technically challenging when working with primary or vivo. Here, we use engineered murine leukemia virus-like particles loaded Cas9-sgRNA ribonucleoproteins (Nanoblades) to induce efficient genome-editing cell lines including human induced pluripotent stem cells, hematopoietic mouse bone-marrow cells. Transgene-free Nanoblades are also...
Calcitonin gene-related peptide (CGRP) is a neuropeptide that coexists with acetylcholine in spinal cord motoneurons. The effects of CGRP on the functional properties nicotinic receptor (AcChoR) were examined by electrophysiological methods. Using whole-cell patch-clamp technique and mouse cell line derived from soleus muscle, we found produces progressive reversible enhancement rapid-decay phase AcChoR desensitization. Single-channel data further show decreases acetylcholine-activated...
Amplification of the mRNA polymerase chain reaction is a very sensitive technique to detect low‐abundance transcripts. We describe in this paper conditions necessary make quantitative. Quantification performed exponential phase amplification process and results are standardized with respect those obtained an exogenous which co‐reverse‐transcribed co‐amplified same as analyzed The primers chosen different exons distinguish fragments from contaminating DNA. Analysis kinetics parameters...
Abstract Background Duchenne muscular dystrophy (DMD) causes severe disability of children and death young men, with an incidence approximately 1/5000 male births. Symptoms appear in early childhood, a diagnosis made mostly around 4 years old, time where the amount muscle damage is already significant, preventing therapeutic interventions that could be more efficient at halting disease progression. In meantime, precise moment which phenotypes arise—even asymptomatically—is still unknown....
Duchenne muscular dystrophy (DMD) affects myofibers and muscle stem cells, causing progressive degeneration repair defects. It was unknown whether dystrophic myoblasts—the effector cells of growth regeneration—are affected. Using transcriptomic, genome-scale metabolic modelling functional analyses, we demonstrate, for the first time, convergent abnormalities in primary mouse human myoblasts. In Dmd mdx myoblasts lacking full-length dystrophin, expression 170 genes significantly altered....
Calpain 3 is a nonlysosomal cysteine protease whose biological functions remain unknown. We previously demonstrated that this altered in limb girdle muscular dystrophy type 2A patients. Preliminary observations suggested its gene subjected to alternative splicing. In paper, we characterize transcriptional and posttranscriptional events leading alterations involving the NS, IS1, IS2 regions and/or calcium binding domains of mouse calpain (capn3). These can be divided into three groups: (i)...
Myf5 is the earliest-known muscle-specific factor to be expressed in vivo and its expression associated with determination of myoblast lineage. In C2 cells, we show by immunocytolocalization that disappears rapidly from cells which differentiation program has been initiated. proliferating myoblasts, levels MyoD detected cell are very heterogeneous. We find some heterogeneity arises a posttranscriptional regulation cycle. Immunoblotting extracts synchronized cultures reveals undergoes...
Patch‐clamp techniques were used to record acetylcholine‐ (ACh) activated single‐channel currents in cell‐attached membrane patches from myotubes of the mouse cell line, C2. The effects phenothiazine derivative chlorpromazine (CPZ) and hallucinogen phencyclidine (PCP) on ACh‐activated properties studied under conditions where both compounds are positively charged (pH 7.2). conductance was unaffected by either CPZ or PCP at concentrations ranging 10 500 nM. 10‐200 nM‐CPZ led shortened mean...
Proximal upstream flanking sequences of the mouse myosin alkali light chain gene encoding MLC1F and MLC3F, α-cardiac actin chicken for α-subunit acetylcholine receptor were linked to bacterial chioramphenicol acetyl transferase (CAT) transfected into primary cultures derived from skeletal muscle or myogenic cell lines. We demonstrate that MLC1F/MLC3F has two functional promoters. In cultures, a 1200 bp sequence exon 1 (MLC1F) 438 2 (MLC3F) direct CAT activity in myotubes, but not myoblasts...
It was recently shown that a long non-coding RNA (lncRNA), we named the 91H (i.e. antisense H19 transcript), is overexpressed in human breast tumours and contributes trans to expression of Insulin-like Growth Factor 2 (IGF2) gene on paternal chromosome. Our preliminary experiments suggested an transcript having similar function may also be conserved mouse. In present work, further characterise mouse and, using genetic complementation approach KO myoblast cells, show ectopic can up-regulate...
Protein delivery from genetically modified skeletal muscle has been reported previously. However, a stable and prolonged secretion was obtained in immunocompromised or newborn animals only. To evaluate the clinical relevance of this approach, we have transduced myoblasts an adult β-glucuronidase-deficient (MPS VII) mouse with retroviral vectors carrying either human β-glucuronidase cDNA murine erythropoietin (Epo) cDNA. The cells were then grafted into tibialis anterior immunocompetent MPS...
Thyroid hormones are among the positive regulators of muscle development in vivo, but little is known about way they work. We demonstrate here that MyoD1, one master genes controlling myogenesis, a target T3. After proliferating C2 myoblasts have been treated with T3 for 15 h, we observed rise MyoD1 expression at both mRNA and protein levels. This first hormonal control gene reported so far. also provide data which suggest nuclear receptor(s) direct role on transcription: 1) cells express...
We have previously suggested that a repeated sequence motif in the upstream region of human cardiac actin gene 'CC.Ar.GG', where Ar is an (A + T)-rich six-base-pair-sequence, may be important muscle-specific expression this [Minty, A. & Kedes, L. (1986) Mol. Cell Biol. 6, 2125-2136]. Here we show binds nuclear protein, and binding abolished by mutating either CC GG dinucleotides or centre. Mutation nucleotides also abolishes transcription-stimulating activity on promoter. A similar has been...
Summry— Myf‐5 and MyoD are the two muscle regulatory factors expressed from myoblast stage to maintain identity promote subsequent differentiation of precursor cells. To get insight into their role we have studied capacity proliferate differentiate myf‐5 myoD null myoblasts in primary cultures passages. Our results indicate that differ wild type (wt) they undergo precocious differentiation: become myogenin‐ troponin T‐positive fail incorporate bromodeoxyuridine (BrdU) under culture...
We have studied the contribution of endogenous production insulin-like growth factor II (IGFII) and muscle regulatory factor, MyoD, to autonomy differentiation in isolated skeletal myoblasts. Inhibition MyoD IGFII gene expression myoblasts mouse myogenic cell line, C2, was achieved by transfection selection stably transfected cells (anti-MyoD anti-IGFII cells) with vectors producing or antisense RNA. observed that inhibiting either has multiple similar consequences. In addition inhibition...
Duchenne muscular dystrophy (DMD) is a devastating X-linked recessive genetic myopathy. DMD physiopathology still not fully understood and prenatal onset suspected but difficult to address. The bone morphogenetic protein 4 (BMP4) critical signaling molecule involved in mesoderm commitment. Human induced pluripotent stem cells (hiPSCs) from healthy individuals human embryonic (hESCs) treated with BMP4 allowed us model the early steps of myogenesis normal contexts. Unexpectedly, 72h following...