RNAi-mediated Stable Silencing of HGPRT Expression in Rabbit Fibroblasts and SCNT Embryo Production / 生物化学与生物物理进展

ABSTRACT

The hypoxanthine-guanine phosphoribosyl transferase (HGPRT) gene mutation is responsible for gouty arthritis, kidney stone, and Lesch-Nyhan Syndrome (LNS). It has been reported that the expression of HGPRT is decreased or even absent in these diseases. Rabbits are an ideal model for studying the pathology of these diseases. Therefore, the development of an HGPRT-knockdown rabbit model will be highly beneficial m such studies. Stable HGPRT-knoekdown transgenie fibroblast lines were generated by transfecting rabbit fibroblasts with RNA interference (RNAi) plasmids. Polymerase chain reaction (PCR) analyses indicated that the average positive rate was 83.3%. The mRNA and protein levels of HGPRT in the transgenic fibroblast lines were significantly lower than that in the control. Transgenic rabbit blastocysts were derived after performing nuclear transfer. The results show that RNAi can be used to stably knock down expression of the HGPRT in rabbit fibroblasts and further improvements in related technologies will facilitate the use of this method for the generation of HGPRT-knockdown rabbits.