期刊
ANDROLOGY
卷 7, 期 5, 页码 644-653出版社
WILEY
DOI: 10.1111/andr.12621
关键词
genetically modified mice; genome editing; sperm maturation
类别
资金
- Bill & Melinda Gates Foundation [OPP1160866] Funding Source: Medline
- Japan Agency for Medical Research and Development [JP18gm5010001] Funding Source: Medline
- Japan Society for the Promotion of Science [JP17H01394, JP18K14612, JP25112007, 20170633] Funding Source: Medline
- Ministry of Education, Culture, Sports, Science and Technology Funding Source: Medline
- NICHD NIH HHS [R01 HD088412, P01 HD087157] Funding Source: Medline
- NIGMS NIH HHS [T32 GM120011] Funding Source: Medline
- Takeda Science Foundation Funding Source: Medline
Background Spermatozoa become competent for fertilization during transit through the epididymis. As spermatozoa from the proximal caudal epididymis can fertilize eggs, proteins from the caput and corpus epididymis are required for sperm maturation. Objectives Microarray analysis identified that more than 17,000 genes are expressed in the epididymis; however, few of these genes demonstrate expression restricted to the epididymis. To analyze epididymis-enriched gene function in vivo, we generated knockout (KO) mutations in nine genes that are abundantly expressed in the caput and corpus region of the epididymis. Materials and methods KO mice were generated using the CRISPR/Cas9 system. The histology of the epididymis was observed with hematoxylin and eosin staining. KO males were caged with wild-type females for 3-6 months to check fertility. Results We generated individual mutant mouse lines having indel mutations in Pate1, Pate2, or Pate3. We also deleted the coding regions of Clpsl2, Epp13, and Rnase13, independently. Finally, the 150 kb region encoding Gm1110, Glb1l2, and Glb1l3 was deleted to generate a triple KO mouse line. Histology of the epididymis and sperm morphology of all KO lines were comparable to control males. The females mated with these KO males delivered pups at comparable numbers as control males. Discussion and conclusion We revealed that nine genes abundantly expressed in the caput and corpus epididymis are dispensable for sperm function and male fecundity. CRISPR/Cas9-mediated KO mice generation accelerates the screening of epididymis-enriched genes for potential functions in reproduction.
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