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Genetics

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JoVE Journal Genetics
Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
 

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Article DOI: 10.3791/56969-v • 11:27 min • August 28th, 2018
August 28th, 2018 •
1, 1, 1
1Department of Biochemistry, Purdue University

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요약
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Targeted genome editing in the model system Danio rerio (zebrafish) has been greatly facilitated by the emergence of CRISPR-based approaches. Herein, we describe a streamlined, robust protocol for generation and identification of CRISPR-derived nonsense alleles that incorporates the heteroduplex mobility assay and identification of mutations using next-generation sequencing.

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CRISPR/Cas9 Gene Knockouts Targeted Genome Editing Zebrafish Biological Role Of A Gene Vertebrate Model System Developmental Processes Genomic Modifications Undergraduates Genomic Targets CRISPR Mutagenesis Zebrafish Embryo Microinjection Modified Alleles Template Specific Oligos Cas9 Protein SgRNA In Vitro Transcription Kit
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