Creation of zebrafish knock‐in reporter lines in the nefma gene by Cas9‐mediated homologous recombination

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Creation of zebrafish knock‐in reporter lines in the nefma gene by Cas9‐mediated homologous recombination

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Eschstruth, Alexis | Schneider-Maunoury, Sylvie | Giudicelli, François

International audience. CRISPR/Cas9-based strategies are widely used for genome editing in many organisms, including zebrafish. Although most applications consist in introducing double strand break (DSB)-induced mutations, it is also possible to use CRISPR/Cas9 to enhance homology directed repair (HDR) at a chosen genomic location to create knock-ins with optimally controlled precision. Here, we describe the use of CRISPR/Cas9-targeted DSB followed by HDR to generate zebrafish transgenic lines where exogenous coding sequences are added in the nefma gene, in frame with the endogenous coding sequence. The resulting knock-in embryos express the added gene (fluorescent reporter or KalTA4 transactivator) specifically in the populations of neurons that express nefma, making them convenient tools for research on these populations.

Langue: en

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