CRISPR/Cas9 mediated genomic deletions protocol in zebrafish
Since its first application for site-directed mutagenesis in 2013, the CRISPR/Cas9 system revolutionized genome engineering. In this protocol, João Amorim, Chiara Perrod and collaborators, lead by José Bessa from i3S, present a validated workflow for the generation of targeted genomic deletions in zebrafish. Here, step-by-step details can be found from design, cloning and synthesis of single-guide RNAs and Cas9 mRNA, followed by microinjection in zebrafish embryos and subsequent genotyping 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.
The article entitled CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish was published in STAR protocols.