title: Targeted gene disruption with engineered zinc-finger nucleases (ZFNs).
abstract: Reverse genetics in Xenopus has been limited to knockdown strategies using antisense morpholino oligonucleotides (MOs). Recently, engineered zinc-finger nucleases have been used to induce targeted mutations resulting in null alleles. Zinc-finger nuclease (ZFN) technology has been adapted to induce null mutations in many systems previously refractory to targeted gene inactivation. Here we provide a general protocol for inducing targeted mutations in Xenopus tropicalis using ZFNs, a method to detect resulting mutations, and the steps to generate homozygous mutant embryos.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA. jyoung528@berkeley.edu
authors.forename: John J
authors.initials: JJ
authors.lastname: Young
authors.forename: Richard M
authors.initials: RM
authors.lastname: Harland
mesh_terms: D000818:Animals; D017849:Animals, Inbred Strains; D006063:Chorionic Gonadotropin / Q000008:administration & dosage; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D003851:Deoxyribonucleases / Q000737:chemistry / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000502:physiology; D005260:Female; D005306:Fertilization; D018095:Germ-Line Mutation; D006720:Homozygote; D054643:INDEL Mutation; D008297:Male; D016296:Mutagenesis; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D012102:Reproductive Control Agents / Q000008:administration & dosage; D059386:Reverse Genetics; D014981:Xenopus / Q000235:genetics*; D016335:Zinc Fingers
chemical_list: D006063:Chorionic Gonadotropin; D011993:Recombinant Fusion Proteins; D012102:Reproductive Control Agents; D003851:Deoxyribonucleases