Disruption of the myostatin gene in porcine primary fibroblasts and embryos using zinc-finger nucleases

Mol Cells. 2014 Apr;37(4):302-6. doi: 10.14348/molcells.2014.2209. Epub 2014 Apr 21.

Abstract

Myostatin represses muscle growth by negatively regulating the number and size of muscle fibers. Myostatin lossof- function can result in the double-muscling phenotype and increased muscle mass. Thus, knockout of myostatin gene could improve the quality of meat from mammals. In the present study, zinc finger nucleases, a useful tool for generating gene knockout animals, were designed to target exon 1 of the myostatin gene. The designed ZFNs were introduced into porcine primary fibroblasts and early implantation embryos via electroporation and microinjection, respectively. Mutations around the ZFNs target site were detected in both primary fibroblasts and blastocysts. The proportion of mutant fibroblast cells and blastocyst was 4.81% and 5.31%, respectively. Thus, ZFNs can be used to knockout myostatin in porcine primary fibroblasts and early implantation embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Deoxyribonucleases / genetics
  • Deoxyribonucleases / metabolism*
  • Exons / genetics
  • Fibroblasts / physiology*
  • Food, Genetically Modified*
  • Gene Knockdown Techniques
  • Meat Products
  • Microinjections
  • Muscles / physiology*
  • Myostatin / genetics*
  • Primary Cell Culture
  • Swine
  • Zinc Fingers / genetics

Substances

  • Myostatin
  • Deoxyribonucleases