Targeted gene knockout in chickens mediated by TALENs

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Chickens / genetics*
  • Chimera
  • Egg Proteins / genetics
  • Female
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques / methods*
  • Genetic Engineering / methods*
  • Genomics / methods
  • Germ Cells / cytology
  • Germ Cells / physiology
  • Germ-Line Mutation
  • Luminescent Proteins / genetics
  • Male
  • Molecular Sequence Data
  • Ovalbumin / genetics*
  • Poultry / genetics*
  • Transcription, Genetic / genetics
  • Transgenes / genetics

Substances

  • Egg Proteins
  • Luminescent Proteins
  • fluorescent protein 583
  • Ovalbumin