sp3111RNAi转基因小鼠模型的评价以及sp3111下调表达对受精及早胚发育的影响

目的 评价sp3111 RNAi转基因小鼠模型的应用价值以及探讨SP3111蛋白降低表达对受精及早期胚胎发育的影响.方法 ①采用Real-Time PCR方法分别检测F1代和F4代转基因RNAi小鼠中sp3111 mRNA的表达变化,并比较2代间干扰效率的差异.②将精子分为实验组（F4代小鼠精子组）和对照组（野生小鼠精子组）,进行体外授精实验,观察受精率及碎裂胚胎的发生率.结果 ①F1代和F4代小鼠中干扰效率的差异无统计学意义（P〉0.05）.②实验组的正常胚胎与碎裂胚胎的形成率分别为34.48%和46.21%,与对照组相比均有显著差异（P〈0.05）.结论 ①RNAi转基因小鼠中RNAi诱导的转录沉默可稳定遗传至F4代.②SP3111下调表达对小鼠受精及早期胚胎发育有明显的影响.

Evaluation of Sp3111 RNAi Transgenic Mice Model and Outcome of in vitro Fertilization with Spermatozoa from Sp3111 RNAi Transgenic Mice

Objective To evaluate application value of Sp3111 RNAi transgenic mice model and to study the function of the sp3111 protein in fertilization and early embryo development. Methods ①real- time quantitative PCR was used to analysis the expression of sp3111 mRNA both in F1 and F4 generation, and comparing the suppression efficiency of RNAi between the two generations.②Sperm samples collected from male Sp3111 RNAi transgenic mice（F4 generation） and from wild type mice as control group followed by IVF. The two groups were observed for the rates of fertilization and embryo fragmentation. Results ① No significant difference was found between F1 and F4 generation in suppression efficiency of RNAi Pf^3/4 0.05.② The rates of normal and fragmented embryos of the experimental group were 34.48% and 46.21%, respectively,significantly different from those of the control group （P〈0.05）. Conclusion ①the suppression efficiency of RNAi in Sp3111 RNAi transgenic mice could be transmitted stably to F4 generation. ②The low expression of sp3111 gene remarkably affected fertilization and early embryo development in mice.