Knockout rats via embryo microinjection of zinc-finger nucleases. Science 2009 Jul 24;325(5939):433
Date
07/25/2009Pubmed ID
19628861Pubmed Central ID
PMC2831805DOI
10.1126/science.1172447Scopus ID
2-s2.0-67749106611 (requires institutional sign-in at Scopus site) 822 CitationsAbstract
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.
Author List
Geurts AM, Cost GJ, Freyvert Y, Zeitler B, Miller JC, Choi VM, Jenkins SS, Wood A, Cui X, Meng X, Vincent A, Lam S, Michalkiewicz M, Schilling R, Foeckler J, Kalloway S, Weiler H, Ménoret S, Anegon I, Davis GD, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Jacob HJ, Buelow RAuthor
Aron Geurts PhD Professor in the Physiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBase Sequence
DNA
Embryo, Mammalian
Endodeoxyribonucleases
Feasibility Studies
Female
Gene Knockout Techniques
Green Fluorescent Proteins
Immunoglobulin M
Male
Microinjections
Molecular Sequence Data
Mutagenesis, Site-Directed
RNA, Messenger
Rats
Zinc Fingers
rab GTP-Binding Proteins
