Homology-independent integration of plasmid DNA into the Zebrafish genome

• Verfasst von: Auer, Thomas [VerfasserIn]
• Verfasst von: Del Bene, Filippo [VerfasserIn]
• Titel: Homology-independent integration of plasmid DNA into the Zebrafish genome
• Verf.angabe: Thomas O. Auer, Filippo Del Bene
• E-Jahr: 2016
• Jahr: 28 July 2016
• Umfang: 21 S.
• Fussnoten: Gesehen am 20.08.2020
• Titel Quelle: Enthalten in: Zebrafish
• Ausgabe Quelle: 2nd ed. 2016
• Ort Quelle: New York, NY : Humana Press, 2016
• Jahr Quelle: 2016
• Band/Heft Quelle: (2016), Seite 31-51
• ISBN Quelle: 978-1-4939-3771-4
• DOI: doi:10.1007/978-1-4939-3771-4_3
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: CRISPR/Cas
• Sach-SW: Genome editing
• Sach-SW: Genome engineering
• Sach-SW: Genome modification
• Sach-SW: Homology-independent repair
• Sach-SW: Site-specific nuclease
• Sach-SW: Targeted transgene integration
• Sach-SW: Zebrafish
• K10plus-PPN: 1727506626

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.