Skip to Main content Skip to Navigation
Theses

Validation in vivo de l'implication de nouveaux gènes impliqués dans le développement musculaire des mammifères

Louise Hélary 1
1 GAMAA - Génomique AniMale, Amélioration, Adaptation
PEIRENE - PEIRENE, INRAE - Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement : USC1061
Abstract : Even if the major actors and transduction pathways of muscle development have been identified, there are still unknown regulatory factors. An in vitro RNAi screening performed on C2C12 myoblastic cells has permitted to identify 20 novel genes potentially implicated in myogenesis. During my thesis, two of these genes were invalidated on mouse model using CRISPR/Cas9 technology in order to confirm their implication in vivo. For the first gene, due to an early lethality occurring in homozygous mutated animals, only heterozygous animals were studied and there was no muscular development anomaly detected. A refined study of earlier stages of embryonic development permitted to show the essential role of the protein in these phases. The study of the second gene, still in progress, seems to confirm in vivo the implication of the gene on the myogenesis. In order to avoid embryonic lethality due to germline invalidation and to observe more rapidly the effects of gene invalidation in muscle, we developed a technique of somatic transgenesis based on RNA interference. Lentivirus containing a shRNA expression cassette was injected directly into the tibialis anterior of mice. We validated this approach on Myostatin gene, a well-known negative regulator of muscle development, showing that the decrease of Myostatin gene expression was associated to an increase of muscle fibers area. The same approach was used with three genes and support the hypothesis of the implication of one of them in muscle development. Thus, this approach allows a rapid “in vivo” screening of in vitro identified genes. Nonetheless, some improvements should be brought on the protocol according to the first results.
Document type :
Theses
Complete list of metadata

https://hal.inrae.fr/tel-03153185
Contributor : Odile Duval <>
Submitted on : Friday, February 26, 2021 - 10:43:58 AM
Last modification on : Saturday, February 27, 2021 - 3:29:59 AM

Identifiers

  • HAL Id : tel-03153185, version 1

Collections

Citation

Louise Hélary. Validation in vivo de l'implication de nouveaux gènes impliqués dans le développement musculaire des mammifères. Sciences du Vivant [q-bio]. Université de Limoges, 2019. Français. ⟨tel-03153185⟩

Share

Metrics

Record views

13