title: CRISPR/Cas9 Promotes Functional Study of Testis Specific X-Linked Gene In Vivo.
abstract: 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.
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Minyan
authors.initials: M
authors.lastname: Li
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Huang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Yuxi
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Xiya
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Puping
authors.initials: P
authors.lastname: Liang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.|Key Laboratory of Reproductive Medicine of Guangdong Province, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: Shaoquan
authors.initials: S
authors.lastname: Zhan
authors.affiliation: Key Laboratory of Reproductive Medicine of Guangdong Province, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: Shanbo
authors.initials: S
authors.lastname: Cao
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Songyang
authors.affiliation: Guangdong Province Key Laboratory of Reproductive Medicine, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.|State Key Laboratory of Biocontrol, Institute of Healthy Aging Research and SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.
authors.forename: Junjiu
authors.initials: J
authors.lastname: Huang
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000502:physiology*; D002460:Cell Line; D005298:Fertility / Q000235:genetics; D015870:Gene Expression; D023061:Gene Order; D018390:Gene Targeting; D050172:Genes, X-Linked; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D023421:Models, Animal; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics; D010641:Phenotype; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D013081:Sperm Motility / Q000235:genetics; D013091:Spermatogenesis / Q000235:genetics; D013094:Spermatozoa / Q000378:metabolism; D013737:Testis / Q000378:metabolism*
chemical_list: D009687:Nuclear Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; C000603461:synaptonemal complex protein 2, mouse