Genome editing to understand neural circuits formation : a novel CRISPR/Cas9-based strategy for conditional mutagenesis and functional study of the role of the meteorin gene family in zebrafish neurodevelopment

Abstract : In recent years, the zebrafish (Danio rerio) has emerged as a powerful model organism to study neuronal circuit development and function. To date, different genome editing technologies allow the generation of constitutive mutant alleles, permitting the study of gene loss-of-function in this vertebrate model. Nevertheless, to assess the role of certain loci it might be required a precise spatiotemporal control of gene inactivation. The rst part of my thesis describes a novel strategy for tissue-specific gene disruption based on the CRISPR/Cas9 and the Gal4/UAS systems. The described technique allows the induction of somatic mutations in genetically labeled tissues, cell clones or single cells, making it possible to follow the effect of gene disruption in vivo via reporter gene expression. The second part of the thesis focuses on the functional analysis of the role of the meteorin gene family during neuronal development and axonal targeting in zebra sh. Meteorin family is conserved among vertebrates and its members have been shown to be involved in neuronal progenitor proliferation and differentiation and axonal elongation, in vitro. We used the zebrafish nervous system as a model to dissect the role of Meteorins during embryonic development, focusing on their potential role as novel guidance molecules. Interestingly, we found that genes belonging to the meteorin family are expressed along the midline of the larval central nervous system and at the floor plate in the hindbrain and spinal cord. We generated CRISPR/Cas9 mutant lines carrying out-of-frame deletions in the coding sequence of each member of the zebrafish meteorin family and we performed a comprehensive analysis of the establishment of axonal projections in the mutants. Our data pointed out that metrns loss-of-function affects the earliest process of axonal development, demonstrating a crucial role in the process of axonal outgrowth for this new family of evolutionary conserved guidance molecules.
Document type :
Theses
Complete list of metadatas

Cited literature [407 references]  Display  Hide  Download

https://tel.archives-ouvertes.fr/tel-02301987
Contributor : Abes Star <>
Submitted on : Tuesday, October 1, 2019 - 1:02:30 AM
Last modification on : Wednesday, October 2, 2019 - 1:28:07 AM

File

2017PA066269.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-02301987, version 1

Citation

Flavia de Santis. Genome editing to understand neural circuits formation : a novel CRISPR/Cas9-based strategy for conditional mutagenesis and functional study of the role of the meteorin gene family in zebrafish neurodevelopment. Neurons and Cognition [q-bio.NC]. Université Pierre et Marie Curie - Paris VI, 2017. English. ⟨NNT : 2017PA066269⟩. ⟨tel-02301987⟩

Share

Metrics

Record views

32

Files downloads

10