The improvement of microinsemination techniques and the production of transgenic offspring with GS cell-derived spermatids in the rats

本研究的最终目的是通过改进微授精技术，通过转染的生殖干细胞（GSCs）和胚胎干细胞（ESCs）产生大鼠后代，建立生产基因敲除大鼠的生殖技术。获得的初步结果是，将构建的GSCs进行慢病毒转染和圆形精细胞注射，成功培育出表达GFP基因的转基因大鼠。用电穿孔法转染建立的ESCs，通过囊胚注射获得表达Kusabira-Orange基因的转基因大鼠。这是首次发现GSCs和ESCs对转基因后代的产生有促进作用。

The ultimate aim of this study is to establish reproductive technology for production of knock-out rats, by improving microinsemination techniques and producing rat offspring via transfected germ stem cells (GSCs) and embryonic stem cells (ESCs). As the primary result obtained, transgenic rats expressing GFP gene were successfully produced, when the established GSCs were subjected to lentiviral transfection and round spermatid injection. Transgenic rats expressing Kusabira-Orange gene were also produced via blastocyst injection of ESCs established and transfected by electroporation. It was shown for the first time that the GSCs and ESCs can contribute to the production of transgenic offspring in the rats.