Generation of cloned transgenic cats expressing red fluorescence protein.
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.
Yin
Xi Jun
XJ
Department of Animal Science and Technology, Sunchon National University, Sunchon 540-742, South Korea.
Lee
Hyo Sang
HS
Yu
Xian Feng
XF
Choi
Eugene
E
Koo
Bon Chul
BC
Kwon
Mo Sun
MS
Lee
Young S
YS
Cho
Su Jin
SJ
Jin
Guang Zhen
GZ
Kim
Lyoung Hyo
LH
Shin
Hyoung Doo
HD
Kim
Teoan
T
Kim
Nam Hyung
NH
Kong
Il Keun
IK
D000818:Animals; D030801:Animals, Genetically Modified; D002415:Cats / Q000235:genetics*; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D004622:Embryo, Mammalian; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D014018:Tissue Distribution; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein