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This Article Full Text Full Text (PDF) All Versions of this Article: 78/3/425    most recent biolreprod.107.065185v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Yin, X. J. Articles by Kong, I. K. Search for Related Content PubMed PubMed Citation Articles by Yin, X. J. Articles by Kong, I. K. Agricola Articles by Yin, X. J. Articles by Kong, I. K.

Generation of Cloned Transgenic Cats Expressing Red Fluorescence Protein¹

Department of Animal Science and Technology,³ Sunchon National University, Sunchon 540-742, South Korea Department of Animal Sciences,⁷ Chungbuk National University, Cheongju 361-763, South Korea Department of Physiology,⁵ Catholic University of Daegu School of Medicine, Daegu 705-718, South Korea Division of Applied Life Science,⁴ Gyeongsang National University, Jinju 660-701, South Korea Department of Genetic Epidemiology,⁶ SNP Genetics Inc., Seoul 153-801, South Korea Institute of Agriculture and Life Science,⁸ Gyeongsang National University, Jinju 660-701, South Korea

ABSTRACT

A method for engineering and producing genetically modified cats is important for generating biomedical models of human diseases. Here we describe the use of somatic cell nuclear transfer to produce cloned transgenic cats that systemically express red fluorescent protein. Immature oocytes were collected from superovulating cat ovaries. Donor fibroblasts were obtained from an ear skin biopsy of a white male Turkish Angora cat, cultured for one to two passages, and subjected to transduction with a retrovirus vector designed to transfer and express the red fluorescent protein (RFP) gene. A total of 176 RFP cloned embryos were transferred into 11 surrogate mothers (mean = 16 ± 7.5 per recipient). Three surrogate mothers were successfully impregnated (27.3%) and delivered two liveborn and one stillborn kitten at 65 to 66 days of gestation. Analysis of nine feline-specific microsatellite loci confirmed that the cloned cats were genetically identical to the donor cat. Presence of the RFP gene in the transgenic cat genome was confirmed by PCR and Southern blot analyses. Whole-body red fluorescence was detected 60 days after birth in the liveborn transgenic (TG) cat but not in the surrogate mother cat. Red fluorescence was detected in tissue samples, including hair, muscle, brain, heart, liver, kidney, spleen, bronchus, lung, stomach, intestine, tongue, and even excrement of the stillborn TG cat. These results suggest that this nuclear transfer procedure using genetically modified somatic cells could be useful for the efficient production of transgenic cats.

assisted reproductive technology, cat, cloned, cloned animal, developmental biology, red fluorescence protein, somatic cell nuclear transfer, transgenic, transgenic animal

¹Supported by a grant from the Korea Science and Engineering Foundation (KOSEF; grant M10525010001-05N2501-00110), funded by the Korean government (MOST). E.C., Y.S.L., S.J.C., and G.Z.J. were supported by scholarships from the Post BK21 Program.

Correspondence: ²Il Keun Kong, Division of Applied Life Science, Gyeongsang National University, Jinju 660-701, GyeongNam Province, South Korea. FAX: 82 55 756 7171; e-mail: ikong{at}gnu.kr