Homology-independent integration of plasmid DNA into the Zebrafish genome
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 allo...
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| Main Authors: | , |
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| Format: | Chapter/Article |
| Language: | English |
| Published: |
28 July 2016
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| In: |
Zebrafish
Year: 2016, Pages: 31-51 |
| DOI: | 10.1007/978-1-4939-3771-4_3 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1007/978-1-4939-3771-4_3
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| Author Notes: | Thomas O. Auer, Filippo Del Bene |
| Summary: | 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. |
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| Item Description: | Gesehen am 20.08.2020 |
| Physical Description: | Online Resource |
| ISBN: | 9781493937714 |
| DOI: | 10.1007/978-1-4939-3771-4_3 |