Genome engineering uncovers 54 evolutionarily conserved and testis-enriched genes that are not required for male fertility in mice |
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Author: | Miyata, Haruhiko1; Castaneda, Julio M.2,3,4; Fujihara, Yoshitaka1; |
Organizations: |
1Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 5650871, Japan 2Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030 3Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030
4Center for Reproductive Medicine, Baylor College of Medicine, Houston, TX 77030
5Center for Drug Discovery, Baylor College of Medicine, Houston, TX 77030 6Immunology Frontier Research Center, Osaka University, Suita, Osaka 5650871, Japan 7Graduate School of Medicine, Osaka University, Suita, Osaka 5650871, Japan 8Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 5650871, Japan 9Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030 10Faculty of Biochemistry and Molecular Medicine, University of Oulu, FI-90014, Oulu, Finland 11Wellcome Trust Sanger Institute, Hinxton CB10 1SA, United Kingdom 12Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030 13Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030 |
Format: | article |
Version: | published version |
Access: | open |
Online Access: | PDF Full Text (PDF, 0.9 MB) |
Persistent link: | http://urn.fi/urn:nbn:fi-fe2020050825755 |
Language: | English |
Published: |
National Academy of Sciences of the United States of America,
2016
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Publish Date: | 2020-05-08 |
Description: |
AbstractGene-expression analysis studies from Schultz et al. estimate that more than 2,300 genes in the mouse genome are expressed predominantly in the male germ line. As of their 2003 publication [Schultz N, Hamra FK, Garbers DL (2003) Proc Natl Acad Sci USA 100(21):12201–12206], the functions of the majority of these testis-enriched genes during spermatogenesis and fertilization were largely unknown. Since the study by Schultz et al., functional analysis of hundreds of reproductive-tract–enriched genes have been performed, but there remain many testis-enriched genes for which their relevance to reproduction remain unexplored or unreported. Historically, a gene knockout is the “gold standard” to determine whether a gene’s function is essential in vivo. Although knockout mice without apparent phenotypes are rarely published, these knockout mouse lines and their phenotypic information need to be shared to prevent redundant experiments. Herein, we used bioinformatic and experimental approaches to uncover mouse testis-enriched genes that are evolutionarily conserved in humans. We then used gene-disruption approaches, including Knockout Mouse Project resources (targeting vectors and mice) and CRISPR/Cas9, to mutate and quickly analyze the fertility of these mutant mice. We discovered that 54 mutant mouse lines were fertile. Thus, despite evolutionary conservation of these genes in vertebrates and in some cases in all eukaryotes, our results indicate that these genes are not individually essential for male mouse fertility. Our phenotypic data are highly relevant in this fiscally tight funding period and postgenomic age when large numbers of genomes are being analyzed for disease association, and will prevent unnecessary expenditures and duplications of effort by others. see all
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Series: |
Proceedings of the National Academy of Sciences of the United States of America |
ISSN: | 0027-8424 |
ISSN-E: | 1091-6490 |
ISSN-L: | 0027-8424 |
Volume: | 113 |
Issue: | 28 |
Pages: | 7704 - 7710 |
DOI: | 10.1073/pnas.1608458113 |
OADOI: | https://oadoi.org/10.1073/pnas.1608458113 |
Type of Publication: |
A1 Journal article – refereed |
Field of Science: |
1182 Biochemistry, cell and molecular biology |
Subjects: | |
Funding: |
We thank the Biotechnology Research and Development (nonprofit organization) and Department of Experimental Genome Research for the generation of the mutant mice; Dr. John Nelson for critical review of the manuscript; and Shirley Baker for manuscript editing. This work as supported by MEXT Grants 26830056 (to H.M.), 15H05573 (to Y.F.), 15K14366 (to Y.F.), 15K14367 (to A.I.), 15K06999 (to D.K.), 15K18387 (to M.M.), 15J04519 (to A.O.), 25112007 (toM.I.), 25250014 (toM.I.), and 15K21737 (toM.I.); the Takeda Science Foundation Grant (to Y.F. and M.I.);Wellcome Trust Grants 079643 and 098051 (to R.R.-S.); National Institutes of Health (KOMP) Awards U01-HG004080 (to R.R.-S.); NIH U01 Grant HD060496 (to M.M.M.); the Osaka University International Joint Research Promotion Program; Baylor College of Medicine Training Grant 5T32HD007165-35 (to J.M.C. and D.R.A.); and the Academy of Finland and the Sigrid Juselius Foundation (R.P.-H.). |
Dataset Reference: |
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1608458113/-/DCSupplemental. |
http://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1608458113/-/DCSupplemental |
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Copyright information: |
© The Authors. Freely available online through the PNAS open access option https://doi.org/10.1073/pnas.1608458113. |