CRISPR/Cas9-mediated genome editing reveals 30 testis-enriched genes dispensable for male fertility in mice†

Yonggang Lu; Seiya Oura; Takafumi Matsumura; Asami Oji; Nobuyuki Sakurai; Yoshitaka Fujihara; Keisuke Shimada; Haruhiko Miyata; Tomohiro Tobita; Taichi Noda; Julio M Castaneda; Daiji Kiyozumi; Qian Zhang; Tamara Larasati; Samantha A M Young; Mayo Kodani; Caitlin A Huddleston; Matthew J Robertson; Cristian Coarfa; Ayako Isotani; R John Aitken; Masaru Okabe; Martin M Matzuk; Thomas X Garcia; Masahito Ikawa

doi:10.1093/biolre/ioz103

CRISPR/Cas9-mediated genome editing reveals 30 testis-enriched genes dispensable for male fertility in mice†

Biol Reprod. 2019 Aug 1;101(2):501-511. doi: 10.1093/biolre/ioz103.

Authors

Yonggang Lu¹, Seiya Oura^{1

2}, Takafumi Matsumura^{1

2}, Asami Oji^{1

3}, Nobuyuki Sakurai¹, Yoshitaka Fujihara^{1

4}, Keisuke Shimada¹, Haruhiko Miyata¹, Tomohiro Tobita^{1

5}, Taichi Noda¹, Julio M Castaneda¹, Daiji Kiyozumi^{1

6}, Qian Zhang^{1

7}, Tamara Larasati^{1

5}, Samantha A M Young^{1

8}, Mayo Kodani^{1

2}, Caitlin A Huddleston^{9

10}, Matthew J Robertson^{11

12}, Cristian Coarfa¹³, Ayako Isotani¹⁴, R John Aitken^{8

15}, Masaru Okabe¹, Martin M Matzuk^{9

10

11

13

16

17}, Thomas X Garcia^{9

10

18}, Masahito Ikawa^{1

2

5

6

19}

Affiliations

¹ Department of Experimental Genome Research, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
² Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
³ RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo, Japan.
⁴ Department of Bioscience and Genetics, National Cerebral and Cardiovascular Center, Suita, Osaka, Japan.
⁵ Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
⁶ Immunology Frontier Research Center, Osaka University, Osaka, Japan.
⁷ Laboratory Animal Center, Chongqing Medical University, Chongqing, China.
⁸ Priority Research Centre for Reproductive Science, Faculty of Science, The University of Newcastle, Callaghan, New South Wales, Australia.
⁹ Department of Pathology and Immunology, Baylor College of Medicine, Houston, Texas, USA.
¹⁰ Center for Drug Discovery, Baylor College of Medicine, Houston, Texas, USA.
¹¹ Dan L Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, Texas, USA.
¹² Advanced Technology Cores, Baylor College of Medicine, Houston, Texas, USA.
¹³ Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA.
¹⁴ Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, Japan.
¹⁵ Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia.
¹⁶ Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.
¹⁷ Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, Texas, USA.
¹⁸ Department of Biology and Biotechnology, University of Houston-Clear Lake, Houston, Texas, USA.
¹⁹ The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

Abstract

More than 1000 genes are predicted to be predominantly expressed in mouse testis, yet many of them remain unstudied in terms of their roles in spermatogenesis and sperm function and their essentiality in male reproduction. Since individually indispensable factors can provide important implications for the diagnosis of genetically related idiopathic male infertility and may serve as candidate targets for the development of nonhormonal male contraceptives, our laboratories continuously analyze the functions of testis-enriched genes in vivo by generating knockout mouse lines using the CRISPR/Cas9 system. The dispensability of genes in male reproduction is easily determined by examining the fecundity of knockout males. During our large-scale screening of essential factors, we knocked out 30 genes that have a strong bias of expression in the testis and are mostly conserved in mammalian species including human. Fertility tests reveal that the mutant males exhibited normal fecundity, suggesting these genes are individually dispensable for male reproduction. Since such functionally redundant genes are of diminished biological and clinical significance, we believe that it is crucial to disseminate this list of genes, along with their phenotypic information, to the scientific community to avoid unnecessary expenditure of time and research funds and duplication of efforts by other laboratories.

Keywords: CRISPR/Cas9; knockout mice; male infertility; spermatogenesis; testis expression.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems*
Fertility / genetics*
Gene Editing*
Gene Expression Regulation / physiology*
Humans
Infertility, Male / genetics
Male
Mice
Mice, Knockout
Testis / metabolism*
Transcriptome

Abstract

Publication types

MeSH terms

Grants and funding