基因CR43905敲除对果蝇精子发生和生育力无明显影响

Normal spermatogenesis and fertility in CR43905 mutant Drosophila melanogaster

目的:探究睾丸特异表达的非编码基因CR43905在果蝇精子发生和生育力中的生理功能。方法:通过qRT-PCR检测CR43905在果蝇不同组织部位的表达情况;利用CRISPR/Cas9基因编辑技术构建CR43905敲除的果蝇品系并通过Sanger测序验证基因组DNA水平的缺失;进一步通过相差显微镜和免疫荧光染色方法观察和形态学分析果蝇CR43905缺失对精子发生的各个阶段生精细胞形态的影响;通过生育力测试实验检测CR43905缺失对雄蝇生育能力的影响。结果:果蝇CR43905是一个睾丸特异表达的非编码基因;利用CRISPR/Cas9技术成功获得了CR43905突变的果蝇品系;果蝇CR43905缺失对精子发生中各阶段生精细胞和雄性生育能力无明显影响。结论:非编码基因CR43905对于果蝇精子发生和雄性生育力的维持是非必需的。

Objective:To explore the biological function in spermatogenesis and fertility of testis-specific expressed non-coding gene CR43905 in Drosophila melanogaster.Methods:Quantitative real-time PCR(qRT-PCR)was used to detect the expression of CR43905 RNA in different drosophila tissues.The CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9)system and Sanger sequencing were used to generate and verify CR43905 knockout mutation drosophila strain,respectively.Phase-contrast microscope and immunofluorescence were used to observe and analyze the histological variation in the development of spermatogenic cells between CR43905 mutant and wildtype drosophila.Male fertility test was conducted to evaluate the influence of drosophila CR43905 mutation on the fertility.Results:Non-coding gene CR43905 exhibits a testis-specific expression pattern in drosophila.We successfully generated a CR43905 mutant drosophila strain using CRISPR/Cas9 system.We found no significant differences of the different stage spermatogenic cells in testis or male fecundity between CR43905 null mutants and wildtype drosophila.Conclusion:Fly CR43905 is dispensable for spermatogenesis and male fecundity.