Generation of gene-specific mutated rats using zinc-finger nucleases

Methods Mol Biol. 2010;597:211-25. doi: 10.1007/978-1-60327-389-3_15.

Abstract

The genetic dissection of physiological and pathological traits in laboratory model organisms is accelerated by the ability to engineer loss-of-function mutations at investigator-specified loci. This chapter describes the use of zinc-finger nucleases (ZFNs) for the targeted disruption of endogenous rat genes directly in the embryo. ZFNs can specifically disrupt target genes in cultured rat cells and in embryos from inbred and outbred strains, leading to permanently genetically modified animals. This technology allows for the rapid, targeted modification of the rat genome.

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics*
  • Cells, Cultured
  • Deoxyribonucleases, Type II Site-Specific / analysis
  • Deoxyribonucleases, Type II Site-Specific / genetics*
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Embryo Transfer
  • Female
  • Gene Knockout Techniques / methods
  • Genes*
  • Genome
  • Male
  • Protein Engineering / methods
  • RNA, Messenger / genetics
  • Rats / genetics*
  • Transcription, Genetic
  • Zinc Fingers*

Substances

  • RNA, Messenger
  • endodeoxyribonuclease FokI
  • Deoxyribonucleases, Type II Site-Specific