1.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
Asian Journal of Andrology 2022;24(3):266-272
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.
Animals
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CRISPR-Cas Systems/genetics*
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Fertility/genetics*
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Gene Editing
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Humans
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Male
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Mice
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Mice, Knockout
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Testis/metabolism*
2.Cooperation-based sperm clusters mediate sperm oviduct entry and fertilization.
Yongcun QU ; Qi CHEN ; Shanshan GUO ; Chiyuan MA ; Yonggang LU ; Junchao SHI ; Shichao LIU ; Tong ZHOU ; Taichi NODA ; Jingjing QIAN ; Liwen ZHANG ; Xili ZHU ; Xiaohua LEI ; Yujing CAO ; Wei LI ; Wei LI ; Nicolas PLACHTA ; Martin M MATZUK ; Masahito IKAWA ; Enkui DUAN ; Ying ZHANG ; Hongmei WANG
Protein & Cell 2021;12(10):810-817
Animals
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Antigens, Surface/genetics*
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Cell Communication/genetics*
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Copulation/physiology*
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Fallopian Tubes/metabolism*
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Female
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Fertilization/genetics*
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GPI-Linked Proteins/genetics*
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Gene Expression Regulation
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Genes, Reporter
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Green Fluorescent Proteins/metabolism*
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Litter Size
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Luminescent Proteins/metabolism*
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Mitochondria/metabolism*
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Reproduction/genetics*
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Signal Transduction
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Sperm Count
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Sperm Motility/genetics*
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Spermatozoa/metabolism*
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Uterus/metabolism*