Thousands of genes are involved in spermatogenesis, however, the functional roles of most these genes for male fertility remain to be discovered. This research focused to explore the function of evolutionarily conserved and testis-specific expressed gene 4930524B15Rik, which is known as C5orf47 in human. We generated 4930524B15Rik knockout mice by CRISPR/Cas9 technology and found 4930524B15Rik−/− mice were fertile. Furthermore, no averted abnormalities were observed in testis morphology, epididymal sperm contents and sperm morphology in 4930524B15Rik knockout mice. Subsequently, histological analysis of testicular tissue revealed intact structure of seminiferous tubules along with the presence of all types of germ cells in 4930524B15Rik−/− mice similar to wild type. Additionally, cytological analysis of spermatocytes displayed no significant differences in the prophase I progression of meiosis, further indicating that 4930524B15Rik have no essential function in mammalian spermatogenesis. Altogether, these results indicated that 4930524B15Rik is dispensable for fertility of male mice and these findings will help researchers to avoid future research overlap and to focus on genes that are crucial for spermatogenesis and reproduction.
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This work was supported by the National Key Research and Developmental Program of China (2018YFC1004700, 2018YFC1003403 and 2016YFC1000600), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000), the National Natural Science Foundation of China (31890780, 31630050 and 31871514), Major Program of Development Foundation of Hefei Centre for Physical Science and Technology (2018ZYFX005), the Fundamental Research Funds for the Central Universities (YD2070002006).
Conflict of interest
All authors declare that no conflict of interest exists.
All the experiments and examination on laboratory animals were directed by the institutional rules of Institutional Animal Care Committee of University of Science and Technology of China with the approval number USTCACUC1301021.
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Fig. S1 mRNA sequencing and real-time PCR. (A) Representative Sanger sequencing chromatograms at cDNA levels confirmed the insertion of nucleotide of 930524B15Rik in knockout mice. Red arrowhead above the chromatograms cDNA sequence show the insertion. (B) Real-time PCR revealed significant decrease of 4930524B15Rik mRNA in 4930524B15Rik-/- testes. (TIF 759 kb)
Fig. S2 4930524B15Rik-/- sperm morphology and motility. (A) Sperm morphology from 10-week-old 4930524B15Rik+/+ and 4930524B15Rik-/- mice. Scale bars, 50 μm. (B) The ratio of normal or abnormal sperm from 10-week-old 4930524B15Rik+/+ and 4930524B15Rik-/- mice. (C) The ratio of average rate of motile sperm in 4930524B15Rik+/+ and 4930524B15Rik-/- mice. (D) The ratio of progressive motile sperm from 4930524B15Rik+/+ and 4930524B15Rik-/- mice. (TIF 6626 kb)
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Khan, R., Ye, J., Yousaf, A. et al. Evolutionarily conserved and testis-specific gene, 4930524B15Rik, is not essential for mouse spermatogenesis and fertility. Mol Biol Rep (2020). https://doi.org/10.1007/s11033-020-05595-0
- Testis-specific gene