Effects of m6A reader Ythdf1 during spermatogenesis
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摘要:
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。
Abstract:
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.
