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Title: Homology-Independent Integration of Plasmid DNA into the Zebrafish Genome. Author: Auer TO, Del Bene F. Journal: Methods Mol Biol; 2016; 1451():31-51. PubMed ID: 27464799. 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.[Abstract] [Full Text] [Related] [New Search]