Abstract
Somatic cell nuclear transfer (SCNT) has been considered as an important method for the production of genetically identical animals and propagation of valuable genotypes. A 3-year-old dog with an outstanding talent for sniffing a cancer was used as a donor dog for SCNT. The donor dog cannot get pregnant by natural fertilization after she suffered pyometra leading to the removal of her reproductive tract. Thus, the aim of this study was to preserve the cancer sniffing dog with excellent genetic background by SCNT. Skin fibroblast from a piece of ear tissue was isolated and cultured. For SCNT, in vivo matured dog oocytes were collected by flushing the oviducts of estrus females. The oocytes were enucleated, microinjected with a donor cell, and fused by electric stimulation. One hundred thirty out of 163, reconstructed embryos (79.8%) were successfully fused. The cloned embryos were chemically activated and transferred into the uterine tube of 7 naturally estrus-synchronized surrogates as with previous report (Lee et al. 2005 Nature). One among them became pregnant, maintained to term, and delivered 4 cloned live offspring (3.1%, live birth/transferred embryos), weighting 450, 410, 430, and 420 g by Caesarean section. The parentage analysis was performed and all cloned pups were genetically identical to the donor dog, and their mitochondrial DNA was originated from their oocyte donor dogs. The present results demonstrated the successful production of four cloned cancer sniffing dogs by SCNT. To date, all cloned puppies have been healthy, growing without apparent abnormalities. Cloned dogs will be trained for cancer sniffing and will be able to provide potential scientific evidence toward the cancer detecting mechanism. This study was financially supported by KOSEF (grant # M10625030005-08N250300510), SNU foundation (Benefactor; RNL BIO), and the Korean MEST, through the BK21 program for Veterinary Science.
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