Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair

Sequence-specific nucleases like TALENs and the CRISPR/Cas9 system have greatly expanded the genome editing possibilities in model organisms such as zebrafish. Both systems have recently been used to create knock-out alleles with great efficiency, and TALENs have also been successfully employed in knock-in of DNA cassettes at defined loci via homologous recombination (HR). Here we report CRISPR/Cas9-mediated knock-in of DNA cassettes into the zebrafish genome at a very high rate by homology-independent double-strand break (DSB) repair pathways. After co-injection of a donor plasmid with a short guide RNA (sgRNA) and Cas9 nuclease mRNA, concurrent cleavage of donor plasmid DNA and the selected chromosomal…

• IE
  Infrastructures en Biologie Santé et Agronomie
• BI
  Boehringer Ingelheim Fonds
• AN
  Agence Nationale de la Recherche
• IN
  Institut National de la Santé et de la Recherche Médicale
• CN
  Centre National de la Recherche Scientifique

  Award: UMR3215