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Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12716-21. doi: 10.1073/pnas.1410555111. Epub 2014 Aug 19.

Targeted gene knockout in chickens mediated by TALENs.

Author information

1
Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea; Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do 232-916, Korea; and.
2
Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea;
3
Department of Chemistry, College of Natural Science, Seoul National University, Seoul 151-921, Korea.
4
Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea; jaehan@snu.ac.kr.

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.

KEYWORDS:

egg protein; genetic modification; germ-line chimera; poultry; programmable genome editing

PMID:
25139993
PMCID:
PMC4156757
DOI:
10.1073/pnas.1410555111
[Indexed for MEDLINE]
Free PMC Article

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