PUBLICATION
Homology-Independent Integration of Plasmid DNA into the Zebrafish Genome
- Authors
- Auer, T.O., Del Bene, F.
- ID
- ZDB-PUB-160729-27
- 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.
Citation
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.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping