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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kim SC, Skowron PM, Szybalski W (1996) Structural requirements for FokI-DNA interaction and oligodeoxyribonucleotide-instructed cleavage. J Mol Biol 258:638–649
Klug A (2005) Towards therapeutic applications of engineered zinc finger proteins. FEBS Lett 579:892–894
Pabo CO, Peisach E, Grant RA (2001) Design and selection of novel Cys2His2 zinc finger proteins. Annu Rev Biochem 70:313–340
Porteus MH, Carroll D (2005) Gene targeting using zinc finger nucleases. Nat Biotechnol 23:967–973
Urnov FD, Miller JC, Lee YL, Beausejour CM, Rock JM, Augustus S, Jamieson AC, Porteus MH, Gregory PD, Holmes MC (2005) Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 435:646–651
Wyman C, Kanaar R (2006) DNA double-strand break repair: all’s well that ends well. Annu Rev Genet 40:363–383
Lieber MR (2008) The mechanism of human nonhomologous DNA end joining. J Biol Chem 283:1–5
Bibikova M, Golic M, Golic KG, Carroll D (2002) Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases. Genetics 161:1169–1175
Carroll D (2008) Progress and prospects: zinc-finger nucleases as gene therapy agents. Gene Ther 15:1463–1468
Geurts AM, Cost FC, Miller JC, Freyvert Y, Zeitler B, Choi VM, Jenkins SS, Wood A, Cui X, Meng X, Vincent A, Lam S, DeKelver RC, Michalkiewicz M, Schilling R, Foeckler J, Kalloway S, Weiler H, Menoret S, Anegon I, Davis GD, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Jacob HJ, Buelow R (2009) Knockout rats via embryo microinjection of zinc finger nucleases. Science 325:433
Si-Hoe SL, Murphy D (1999) Production of transgenic rodents by the microinjection of cloned DNA into fertilized one-celled eggs. Methods Mol Biol 97:61–100
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Geurts, A.M. et al. (2010). Generation of Gene-Specific Mutated Rats Using Zinc-Finger Nucleases. In: Anegon, I. (eds) Rat Genomics. Methods in Molecular Biology, vol 597. Humana Press. https://doi.org/10.1007/978-1-60327-389-3_15
Download citation
DOI: https://doi.org/10.1007/978-1-60327-389-3_15
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60327-388-6
Online ISBN: 978-1-60327-389-3
eBook Packages: Springer Protocols