CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish

STAR Protocols

Renata Bordeira-Carriço¹  Chiara Perrod¹  João Pedro Amorim²  Ana Gali-Macedo²  José Bessa² 

[1] IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal;i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal;

关键词: CRISPR;    Genetics;    Genomics;    Model Organisms;   

DOI  :  

来源: DOAJ

【 摘 要 】

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).

【 授权许可】

Unknown