CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice
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 us...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wolters Kluwer Medknow Publications
2022-01-01
|
| Series: | Asian Journal of Andrology |
| Subjects: | |
| Online Access: | http://www.ajandrology.com/article.asp?issn=1008-682X;year=2022;volume=24;issue=3;spage=266;epage=272;aulast=Oyama |
| _version_ | 1828336169361866752 |
|---|---|
| author | Yuki Oyama Haruhiko Miyata Keisuke Shimada Yoshitaka Fujihara Keizo Tokuhiro Thomas X Garcia Martin M Matzuk Masahito Ikawa |
| author_facet | Yuki Oyama Haruhiko Miyata Keisuke Shimada Yoshitaka Fujihara Keizo Tokuhiro Thomas X Garcia Martin M Matzuk Masahito Ikawa |
| author_sort | Yuki Oyama |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T21:57:27Z |
| format | Article |
| id | doaj.art-7f88ba8a95df47dfb5206782c6123575 |
| institution | Directory Open Access Journal |
| issn | 1008-682X 1745-7262 |
| language | English |
| last_indexed | 2024-04-13T21:57:27Z |
| publishDate | 2022-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Asian Journal of Andrology |
| spelling | doaj.art-7f88ba8a95df47dfb5206782c61235752022-12-22T02:28:12ZengWolters Kluwer Medknow PublicationsAsian Journal of Andrology1008-682X1745-72622022-01-0124326627210.4103/aja.aja_63_21CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in miceYuki OyamaHaruhiko MiyataKeisuke ShimadaYoshitaka FujiharaKeizo TokuhiroThomas X GarciaMartin M MatzukMasahito IkawaGene 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.http://www.ajandrology.com/article.asp?issn=1008-682X;year=2022;volume=24;issue=3;spage=266;epage=272;aulast=Oyamacrispr/cas9; knockout mice; male infertility; spermatozoa; testis |
| spellingShingle | Yuki Oyama Haruhiko Miyata Keisuke Shimada Yoshitaka Fujihara Keizo Tokuhiro Thomas X Garcia Martin M Matzuk Masahito Ikawa CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice Asian Journal of Andrology crispr/cas9; knockout mice; male infertility; spermatozoa; testis |
| title | CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice |
| title_full | CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice |
| title_fullStr | CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice |
| title_full_unstemmed | CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice |
| title_short | CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice |
| title_sort | crispr cas9 mediated genome editing reveals 12 testis enriched genes dispensable for male fertility in mice |
| topic | crispr/cas9; knockout mice; male infertility; spermatozoa; testis |
| url | http://www.ajandrology.com/article.asp?issn=1008-682X;year=2022;volume=24;issue=3;spage=266;epage=272;aulast=Oyama |
| work_keys_str_mv | AT yukioyama crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT haruhikomiyata crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT keisukeshimada crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT yoshitakafujihara crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT keizotokuhiro crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT thomasxgarcia crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT martinmmatzuk crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice AT masahitoikawa crisprcas9mediatedgenomeeditingreveals12testisenrichedgenesdispensableformalefertilityinmice |