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JoVE Journal
Genetics

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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

Chapters

Summary

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.

Tags

Keywords: CRISPR/Cas9 Gene Knockout Danio Rerio Zebrafish Targeted Genome Editing In Vitro Transcription SgRNA Ammonium Acetate Precipitation Ethanol Precipitation RNA Purification
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