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The CRISPR and in vivo recombination: news tools for studying sex determination in the Medaka fish

Abstract : In medaka, dmrt1bY, the duplicated copy on the Y-chromosome of dmrt1a, became the master regulator of male development after acquiring an upstream position in the sex-determining cascade. Remarkably this evolutionary novelty, requiring a rewiring of the regulatory network, was brought about by co-optation of pre-existing cis-regulatory elements contributed by transposable elements. To another level of regulation, differential gene expression of the two dmrt1 co-orthologs seems to be also mediated by post-transcriptional regulation through a highly conserved cis-regulatory motif that directs differential gonadal synexpression of Dmrt1 transcripts during gonad development. For a detailed analysis of expression, regulation and function of dmrt1 and other gonadal genes, we generated several transgenic fluorescent reporter lines (BAC clones). Further on, using genome editing (CRISPR-Cas9) as well as conditional in vivo knock-in (KI), we found clear evidences that the specific expression pattern of dmrt1bY is controlled by (i) differential cross-regulations of dmrt1a, dmrt1bY and a HMG-box gene, (ii) specific stabilization and translational control of dmrt1 transcripts via an RNA-binding protein. Into that context, we will give special emphasis on the CRISPR method we employed for medaka fish as well as the conditional in vivo recombination. We will share our experience and discuss our results regarding to the generation and the use of such fish.
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Conference papers
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https://hal.inrae.fr/hal-02801182
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Submitted on : Friday, June 5, 2020 - 8:29:58 PM
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  • HAL Id : hal-02801182, version 1
  • PRODINRA : 284728

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Amaury Herpin. The CRISPR and in vivo recombination: news tools for studying sex determination in the Medaka fish. CRISPR/CAS9 pour modèles aquatiques. Séminaire et table ronde, Nov 2014, Rennes, France. ⟨hal-02801182⟩

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