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Genesis. 2019 Sep 30:e23340. doi: 10.1002/dvg.23340. [Epub ahead of print]

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

Author information

1
Sorbonne Université, CNRS UMR7622, Inserm U1156, Institut de Biologie Paris-Seine (IBPS) - Laboratoire de Biologie du développement, Paris, France.

Abstract

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.

KEYWORDS:

CRISPR/Cas9; genome editing; knock-in; nefma; zebrafish

PMID:
31571409
DOI:
10.1002/dvg.23340

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