|ZFIN ID: ZDB-PUB-160729-27|
Homology-Independent Integration of Plasmid DNA into the Zebrafish Genome
Auer, T.O., Del Bene, F.
|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|
|PubMed:||27464799 Full text @ Meth. Mol. Biol.|
Auer, T.O., Del Bene, F. (2016) Homology-Independent Integration of Plasmid DNA into the Zebrafish Genome. Methods in molecular biology (Clifton, N.J.). 1451:31-51.
ABSTRACTTargeting 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.
ADDITIONAL INFORMATION No data available