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.
Date: 2019
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
MeSH Terms:
  • Animals
  • Animals, Genetically Modified/genetics
  • CRISPR-Cas Systems/genetics
  • Gene Editing/methods
  • Gene Knock-In Techniques/methods*
  • Gene Targeting/methods
  • Genetic Engineering/methods*
  • Genome/genetics
  • Homologous Recombination/genetics
  • Intermediate Filaments/genetics
  • RNA, Guide/genetics
  • Zebrafish/genetics
PubMed: 31571409 Full text @ Genesis
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.