Publication Date
1-1-2022
Journal
Asian Journal of Andrology
DOI
10.4103/aja.aja_63_21
PMID
34290169
PMCID
PMC9226692
PubMedCentral® Posted Date
7-16-2021
PubMedCentral® Full Text Version
Post-print
Published Open-Access
yes
Keywords
Animals, CRISPR-Cas Systems, Fertility, Gene Editing, Humans, Male, Mice, Mice, Knockout, Testis, CRISPR/Cas9, knockout mice, male infertility, spermatozoa, testis
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.
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Immunology of Infectious Disease Commons, Immunopathology Commons, Medical Immunology Commons, Pathology Commons