Generation of Targeted Genomic Deletions Through CRISPR/Cas System in Zebrafish

Abstract

Using TALEN or CRISPR/Cas system to induce small indels into coding sequences has been implicated in broad applications for reverse genetic studies of many organisms including zebrafish. However, complete deletion of a large gene or noncoding gene(s) or removing a large genomic fragment spanning several genes or other chromosomal elements is preferred in various cases, as well as inducing chromosomal inversions. Here, we describe the detailed protocols for the generation of chromosomal deletion mutations mediated by Cas9 and a pair of gRNAs and the evaluation for the efficiencies in F0 founder fish and of germline transmission.

Keywords: CRISPR/Cas; Chromosomal deletion; Gene disruption; Gene targeting; Genome manipulation; TALEN; Zebrafish.