Production of gene Knock-down animal by introducing RNAi into spermatogonial stem cells.

为了评估RNAi介导的基因敲除是否可以应用于生殖系干细胞，将EGFP敲除的RNAi构建物转导到EGFP转基因小鼠的生殖系干细胞中。虽然慢病毒介导的RNAi转导在一定程度上抑制了种系干细胞中EGFP的表达，但这种抑制效果不足以产生敲除小鼠。同时，通过本项目开发的RNAi技术，我们发现DNA甲基转移酶I对于种系干细胞的体外存活是必不可少的，提示RNAi介导的敲低内源基因是可行的。

To assess whether RNAi mediated gene knock-down could be applied to germline stem cells, RNAi construct for EGFP knock-down was transduced to germline stem cells derived from EGFP transgenic mouse. Although lentivirus mediated RNAi transduction suppressed the EGFP expression to some extent in germline stem cells, the efficiency was not enough to produce the knock-down mouse. Meanwhile, by using RNAi technology developed in this project, it was shown that DNA methyl transferase I is indispensable for the survival of germline stem cells in vitro, suggesting that RNAi mediated knockdown is available for the endogenous genes.