title: Finding of a highly efficient ZFN pair for Aqpep gene functioning in murine zygotes.
abstract: The generation efficiencies of mutation-induced mice when using engineered zinc-finger nucleases (ZFNs) have been generally 10 to 20% of obtained pups in previous studies. The discovery of high-affinity DNA-binding modules can contribute to the generation of various kinds of novel artificial chromatin-targeting tools, such as zinc-finger acetyltransferases, zinc-finger histone kinases and so on, as well as improvement of reported zinc-finger recombinases and zinc-finger methyltransferases. Here, we report a novel ZFN pair that has a highly efficient mutation-induction ability in murine zygotes. The ZFN pair induced mutations in all obtained mice in the target locus, exon 17 of aminopeptidase Q gene, and almost all of the pups had biallelic mutations. This high efficiency was also shown in the plasmid DNA transfected in a cultured human cell line. The induced mutations were inherited normally in the next generation. The zinc-finger modules of this ZFN pair are expected to contribute to the development of novel ZF-attached chromatin-targeting tools.
authors.affiliation: Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Fujii
authors.forename: Asuka
authors.initials: A
authors.lastname: Onuma
authors.forename: Shin
authors.initials: S
authors.lastname: Yoshioka
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nagashima
authors.forename: Koji
authors.initials: K
authors.lastname: Sugiura
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Naito
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D051379:Mice; D008845:Microinjections; D009154:Mutation; D010957:Plasmids; D016335:Zinc Fingers / Q000235:genetics*; D015053:Zygote