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Chapters
- 00:05Title
- 01:09Preparation of CRISPR-reagents for Embryo Microinjection
- 04:03Microinjection of CRISPR-components into Zebrafish Embryos
- 06:12Analysis of Efficiency of Indel Formation Using an HMA
- 08:45Identification and Propagation of Knock-out Lines
- 09:45Results: Zebrafish Embryos Exhibit a Pigment Defect when Injected with an sgRNA Targeting Tyrosinase at the One Cell Stage
- 10:32Conclusion
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.
