|ZFIN ID: ZDB-PUB-191002-4|
Creation of zebrafish knock-in reporter lines in the nefma gene by Cas9-mediated homologous recombination
Eschstruth, A., Schneider-Maunoury, S., Giudicelli, F.
|Source:||Genesis (New York, N.Y. : 2000) 58(1): e23340 (Journal)|
|Registered Authors:||Schneider-Maunoury, Sylvie|
|Keywords:||CRISPR/Cas9, genome editing, knock-in, nefma, zebrafish|
|PubMed:||31571409 Full text @ Genesis|
Eschstruth, A., Schneider-Maunoury, S., Giudicelli, F. (2019) Creation of zebrafish knock-in reporter lines in the nefma gene by Cas9-mediated homologous recombination. Genesis (New York, N.Y. : 2000). 58(1):e23340.
ABSTRACTCRISPR/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.