m6A识别蛋白Ythdf1在精子发生中的作用研究

Effects of m6A reader Ythdf1 during spermatogenesis

目的:构建YTH N6甲基腺苷RNA结合蛋白1(YTH N6-methyladenosine RNA binding protein 1,Ythdf1)的基因敲除小鼠,研究其在小鼠精子发生中的作用。方法:利用CRISPR/Cas9敲除技术及原核注射技术构建Ythdf1基因敲除小鼠模型,通过免疫荧光、HE染色对敲除小鼠睾丸及附睾进行形态学分析,通过交配实验进行生殖力测试,研究Ythdf1基因对雄性小鼠精子发生的影响。结果:RT-PCR结果显示,Ythdf1在1～8周龄雄鼠睾丸中均有表达;成功构建Ythdf1基因敲除小鼠;成年敲除小鼠精子发生、附睾及生育力均无异常。结论:在正常饲养情况下,敲除Ythdf1基因对雄性小鼠精子发生无影响。

Objective:Construct YTH N6-methyladenosine RNA binding protein 1(Ythdf1)knockout mice and study its role during mouse spermatogenesis. Methods:Ythdf1 knockout mice were generated by microinjection of CRISPR/Cas9 into zygote successfully. The role of Ythdf1 in mouse spermatogenesis was detected by immunofluorescence,HE and fertility test. Results:The results of RT-PCR showed that Ythdf1 was expressed in the testis of 1 to 8-week-old male mice. Ythdf1 knockout mice were successfully generated. These knockout mice showed normal spermatogenesis,epididymis morphology and fertility. Conclusion:This study found that deletion of Ythdf1 gene had no effect on spermatogenesis under normal breeding conditions.