title: Generation of hypoxanthine phosphoribosyltransferase gene knockout rabbits by homologous recombination and gene trapping through somatic cell nuclear transfer.
abstract: The rabbit is a common animal model that has been employed in studies on various human disorders, and the generation of genetically modified rabbit lines is highly desirable. Female rabbits have been successfully cloned from cumulus cells, and the somatic cell nuclear transfer (SCNT) technology is well established. The present study generated hypoxanthine phosphoribosyltransferase (HPRT) gene knockout rabbits using recombinant adeno-associated virus-mediated homologous recombination and SCNT. Gene trap strategies were employed to enhance the gene targeting rates. The male and female gene knockout fibroblast cell lines were derived by different strategies. When male HPRT knockout cells were used for SCNT, no live rabbits were obtained. However, when female HPRT(+/-) cells were used for SCNT, live, healthy rabbits were generated. The cloned HPRT(+/-) rabbits were fertile at maturity. We demonstrate a new technique to produce gene-targeted rabbits. This approach may also be used in the genetic manipulation of different genes or in other species.
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Mingru
authors.initials: M
authors.lastname: Yin
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Zhenfu
authors.initials: Z
authors.lastname: Fang
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Kong
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Fengying
authors.initials: F
authors.lastname: Xing
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Xuejin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Laboratory Animal Science, School of Medicine, Shanghai Jiao Tong University, 200025 Shanghai, China.
authors.forename: Shangang
authors.initials: S
authors.lastname: Li
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D005260:Female; D055786:Gene Knockout Techniques / Q000379:methods; D059765:Homologous Recombination / Q000235:genetics*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D008297:Male; D053652:Nuclear Transfer Techniques; D011817:Rabbits
chemical_list: D007041:Hypoxanthine Phosphoribosyltransferase