ZFIN ID: ZDB-PUB-160729-27
Homology-Independent Integration of Plasmid DNA into the Zebrafish Genome
Auer, T.O., Del Bene, F.
Date: 2016
Source: Methods in molecular biology (Clifton, N.J.)   1451: 31-51 (Chapter)
Registered Authors: Auer, Thomas, Del Bene, Filippo
Keywords: CRISPR/Cas, Genome editing, Genome engineering, Genome modification, Homology-independent repair, Site-specific nuclease, Targeted transgene integration, Zebrafish
MeSH Terms:
  • Alleles
  • Animals
  • CRISPR-Cas Systems/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • DNA Breaks, Double-Stranded
  • Genetic Engineering
  • Plasmids/genetics*
  • Zebrafish/genetics*
PubMed: 27464799 Full text @ Meth. Mol. Biol.
Targeting nucleases like zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) system have revolutionized genome-editing possibilities in many model organisms. They allow the generation of loss-of-function alleles by the introduction of double-strand breaks at defined sites within genes, but also more sophisticated genome-editing approaches have become possible. These include the integration of donor plasmid DNA into the genome by homology-independent repair mechanisms after CRISPR/Cas9-mediated cleavage. Here we present a protocol outlining the most important steps to target a genomic site and to integrate a donor plasmid at this defined locus.