Targeted gene knockout in chickens mediated by TALENs

Abstract

Genetically modified animals are used for industrial applications as well as scientific research, and studies on these animals contribute to a better understanding of biological mechanisms. Gene targeting techniques have been developed to edit specific gene loci in the genome, but the conventional strategy of homologous recombination with a gene-targeted vector has low efficiency and many technical complications. Here, we generated specific gene knockout chickens through the use of transcription activator-like effector nuclease (TALEN)-mediated gene targeting. In this study, we accomplished targeted knockout of the ovalbumin (OV) gene in the chicken primordial germ cells, and OV gene mutant offspring were generated through test-cross analysis. TALENs successfully induced nucleotide deletion mutations of ORF shifts, resulting in loss of chicken OV gene function. Our results demonstrate that the TALEN technique used in the chicken primordial germ cell line is a powerful strategy to create specific genome-edited chickens safely for practical applications.genetic modification | programmable genome editing | germ-line chimera | poultry | egg protein

The yin and yang of kidney development and Wilms’ tumors

“…Mice might very well be one of the most difficult model organisms to model Wilms' tumors. However, the recent advances in gene editor technologies, such as zinc finger nucleases, TAL effector nucleases, and CRISPR/Cas9, have opened up genomic engineering possibilities in these species to comparable levels that so far have only been possible in mice (Li et al 2013a,b;Honda et al 2014;Park et al 2014;Yang et al 2014). Maybe the time is ripe to reconsider the best possible experimental model for Wilms' tumor research.…”

Leapfrogging: primordial germ cell transplantation permits recovery of CRISPR/Cas9-induced mutations in essential genes

“…PGCs have been transplanted from cultured early chick extraembryonic tissue and can contribute to the germline in recipients (van de Lavoir et al, 2006;Nakamura et al, 2013). We envision combining this technology with programmable nucleases (Park et al, 2014;Véron et al, 2015) to make leapfrogging possible in this and a number of other species.…”

CRISPR mediated somatic cell genome engineering in the chicken

“…Additionally this technology opens the door to further advances in the genetic manipulations of avian species. Indeed, recent reports of genetically modified chickens using targeted gene knockout using the TALEN technology in isolated chicken Primordial Germ Cells (PGCs) (Park et al, 2014) suggests that the same strategy combined with the easier CRISPR technique may soon be used at a large scale to generate specific genome-edited avian lines.…”