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Mammalian spermatogenesis is a highly regulated multistage process of sperm generation. It is hard to uncover the real function of a testis specific gene  in vitro  since the  in vitro  model is not yet mature. With the development of the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated 9) system, we can now rapidly generate knockout mouse models of testis specific genes to study the process of spermatogenesis  in vivo . SYCP3-like X-linked 2 (SLX2) is a germ cell specific component, which contains a Cor1 domain and belongs to the XLR (X-linked, lymphocyte regulated) family. Previous studies suggested that SLX2 might play an important role in mouse spermatogenesis based on its subcellular localization and interacting proteins. However, the function of SLX2  in vivo  is still elusive. Here, to investigate the functions of SLX2 in spermatogenesis, we disrupted the  Slx2  gene by using the CRISPR/Cas9 system. Since  Slx2  is a testis specific X-linked gene, we obtained knockout male mice in the first generation and accelerated the study process. Compared with wild-type mice,  Slx2  knockout mice have normal testis and epididymis. Histological observation of testes sections showed that  Slx2  knockout affected none of the three main stages of spermatogenesis: mitosis, meiosis and spermiogenesis. In addition, we further confirmed that disruption of  Slx2  did not affect the number of spermatogonial stem cells, meiosis progression or XY body formation by immunofluorescence analysis. As spermatogenesis was normal in  Slx2  knockout mice, these mice were fertile. Taken together, we showed that  Slx2  itself is not an essential gene for mouse spermatogenesis and CRISPR/Cas9 technique could speed up the functional study of testis specific X-linked gene  in vivo .

CRISPR/Cas9 Promotes Functional Study of Testis Specific X-Linked Gene In Vivo