CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish

Summary: Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and...

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Bibliographic Details
Main Authors: João Pedro Amorim, Renata Bordeira-Carriço, Ana Gali-Macedo, Chiara Perrod, José Bessa
Format: Article
Language:English
Published: Elsevier 2020-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166720301957
Description
Summary:Summary: Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and Cas9 mRNA, followed by microinjection in zebrafish embryos and subsequent genotype screening for the establishment of a mutant line. The versatility and efficiency of this pipeline makes the generation of zebrafish models a widely used approach in functional genetics.For complete details on the use and execution of this protocol, please refer to Amorim et al. (2020).
ISSN:2666-1667