Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector.

Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo-cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine- and bovine-cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNbeta-EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P < 0.05) development rates of the intraspecies SCNT porcine or bovine embryos did not differ between transgenic and nontransgenic groups. Expression of EGFP was observed in 100% of blastocysts and mosaicism was not observed. Furthermore, after iSCNT of porcine or bovine donor nuclei into xenogenic ooplasm, patterns of nuclear remodeling in reconstructed embryos were similar. In conclusion, our data demonstrated the feasibility of producing transgenic iSCNT embryos. To our knowledge, this is the first report of transgenic cloned embryo production by iSCNT approach. In the future, this may provide a powerful research tool for studying developmental events in domestic animals and provide marked cell lines for other genetic manipulations.

Uhm SJ ，Gupta MK ，Kim T ，Lee HT ... -  《MOLECULAR REPRODUCTION AND DEVELOPMENT》

Effect of transgene introduction and recloning on efficiency of porcine transgenic cloned embryo production in vitro.

Uhm SJ ，Gupta MK ，Das ZC ，Kim JH ，Park C ，Kim T ，Lee HT ... -  《-》

Production of transgenic canine embryos using interspecies somatic cell nuclear transfer.

Hong SG ，Oh HJ ，Park JE ，Kim MJ ，Kim GA ，Koo OJ ，Jang G ，Lee BC ... -  《-》

Production of nuclear transfer-derived piglets using porcine fetal fibroblasts transfected with the enhanced green fluorescent protein.

Hyun S ，Lee G ，Kim D ，Kim H ，Lee S ，Nam D ，Jeong Y ，Kim S ，Yeom S ，Kang S ，Han J ，Lee B ，Hwang W ... -  《BIOLOGY OF REPRODUCTION》

Production of transgenic cloned piglets from genetically transformed fetal fibroblasts selected by green fluorescent protein.

Lee GS ，Kim HS ，Hyun SH ，Lee SH ，Jeon HY ，Nam DH ，Jeong YW ，Kim S ，Kim JH ，Han JY ，Ahn C ，Kang SK ，Lee BC ，Hwang WS ... -  《THERIOGENOLOGY》