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Proc Natl Acad Sci U S A. 2016 Jul 12;113(28):7704-10. doi: 10.1073/pnas.1608458113. Epub 2016 Jun 29.

Genome engineering uncovers 54 evolutionarily conserved and testis-enriched genes that are not required for male fertility in mice.

Author information

1
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 5650871, Japan;
2
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030; Center for Reproductive Medicine, Baylor College of Medicine, Houston, TX 77030;
3
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; Center for Drug Discovery, Baylor College of Medicine, Houston, TX 77030;
4
Immunology Frontier Research Center, Osaka University, Suita, Osaka 5650871, Japan;
5
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; Center for Reproductive Medicine, Baylor College of Medicine, Houston, TX 77030;
6
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 5650871, Japan; Graduate School of Medicine, Osaka University, Suita, Osaka 5650871, Japan;
7
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 5650871, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 5650871, Japan;
8
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030;
9
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; Center for Reproductive Medicine, Baylor College of Medicine, Houston, TX 77030; Faculty of Biochemistry and Molecular Medicine, University of Oulu, FI-90014, Oulu, Finland;
10
Wellcome Trust Sanger Institute, Hinxton CB10 1SA, United Kingdom;
11
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 5650871, Japan; Immunology Frontier Research Center, Osaka University, Suita, Osaka 5650871, Japan; Graduate School of Medicine, Osaka University, Suita, Osaka 5650871, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 5650871, Japan; ikawa@biken.osaka-u.ac.jp mmatzuk@bcm.edu.
12
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030; Center for Reproductive Medicine, Baylor College of Medicine, Houston, TX 77030; Center for Drug Discovery, Baylor College of Medicine, Houston, TX 77030; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030; Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030 ikawa@biken.osaka-u.ac.jp mmatzuk@bcm.edu.

Abstract

Gene-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.

KEYWORDS:

genetically modified mice; genome editing; spermatozoa

PMID:
27357688
PMCID:
PMC4948324
DOI:
10.1073/pnas.1608458113
[Indexed for MEDLINE]
Free PMC Article

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