Simple methods for generating and detecting locus-specific mutations induced with TALENs in the zebrafish genome.

ABSTRACT: The zebrafish is a powerful experimental system for uncovering gene function in vertebrate organisms. Nevertheless, studies in the zebrafish have been limited by the approaches available for eliminating gene function. Here we present simple and efficient methods for inducing, detecting, and recovering mutations at virtually any locus in the zebrafish. Briefly, double-strand DNA breaks are induced at a locus of interest by synthetic nucleases, called TALENs. Subsequent host repair of the DNA lesions leads to the generation of insertion and deletion mutations at the targeted locus. To detect the induced DNA sequence alterations at targeted loci, genomes are examined using High Resolution Melt Analysis, an efficient and sensitive method for detecting the presence of newly arising sequence pol

SUBMITTER: Dahlem TJ 

PROVIDER: S-EPMC3420959 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature