CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice.

Yuki Oyama; Haruhiko Miyata; Keisuke Shimada; Yoshitaka Fujihara; Keizo Tokuhiro; Thomas X Garcia; Martin M Matzuk; Masahito Ikawa

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CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice.

Author: Yuki OYAMA ¹ ; Haruhiko MIYATA ² ; Keisuke SHIMADA ² ; Yoshitaka FUJIHARA ² ; Keizo TOKUHIRO ³ ; Thomas X GARCIA ⁴ ; Martin M MATZUK ⁴ ; Masahito IKAWA ¹
Author Information

1. Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
2. Department of Experimental Genome Research, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
3. Department of Genome Editing, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka 573-1191, Japan.
4. Center for Drug Discovery, Baylor College of Medicine, Houston, TX 77030, USA.
Publication Type:Journal Article
Keywords: CRISPR/Cas9; knockout mice; male infertility; spermatozoa; testis
MeSH: Animals; CRISPR-Cas Systems/genetics*; Fertility/genetics*; Gene Editing; Humans; Male; Mice; Mice, Knockout; Testis/metabolism*
From: Asian Journal of Andrology 2022;24(3):266-272
CountryChina
Language:English
Abstract: Gene expression analyses suggest that more than 1000-2000 genes are expressed predominantly in mouse and human testes. Although functional analyses of hundreds of these genes have been performed, there are still many testis-enriched genes whose functions remain unexplored. Analyzing gene function using knockout (KO) mice is a powerful tool to discern if the gene of interest is essential for sperm formation, function, and male fertility in vivo. In this study, we generated KO mice for 12 testis-enriched genes, 1700057G04Rik, 4921539E11Rik, 4930558C23Rik, Cby2, Ldhal6b, Rasef, Slc25a2, Slc25a41, Smim8, Smim9, Tmem210, and Tomm20l, using the clustered regularly interspaced short palindromic repeats /CRISPR-associated protein 9 (CRISPR/Cas9) system. We designed two gRNAs for each gene to excise almost all the protein-coding regions to ensure that the deletions in these genes result in a null mutation. Mating tests of KO mice reveal that these 12 genes are not essential for male fertility, at least when individually ablated, and not together with other potentially compensatory paralogous genes. Our results could prevent other laboratories from expending duplicative effort generating KO mice, for which no apparent phenotype exists.