Knockout rats via embryo microinjection of zinc-finger nucleases

Science. 2009 Jul 24;325(5939):433. doi: 10.1126/science.1172447.


The toolbox of rat genetics currently lacks the ability to introduce site-directed, heritable mutations into the genome to create knockout animals. By using engineered zinc-finger nucleases (ZFNs) designed to target an integrated reporter and two endogenous rat genes, Immunoglobulin M (IgM) and Rab38, we demonstrate that a single injection of DNA or messenger RNA encoding ZFNs into the one-cell rat embryo leads to a high frequency of animals carrying 25 to 100% disruption at the target locus. These mutations are faithfully and efficiently transmitted through the germline. Our data demonstrate the feasibility of targeted gene disruption in multiple rat strains within 4 months time, paving the way to a humanized monoclonal antibody platform and additional human disease models.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA
  • Embryo, Mammalian
  • Endodeoxyribonucleases / genetics
  • Endodeoxyribonucleases / metabolism*
  • Feasibility Studies
  • Female
  • Gene Knockout Techniques*
  • Green Fluorescent Proteins
  • Immunoglobulin M / genetics*
  • Male
  • Microinjections*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • RNA, Messenger
  • Rats
  • Zinc Fingers* / genetics
  • rab GTP-Binding Proteins / genetics*


  • Immunoglobulin M
  • RNA, Messenger
  • Green Fluorescent Proteins
  • DNA
  • Endodeoxyribonucleases
  • Rab38 protein, rat
  • rab GTP-Binding Proteins