Restore natural fertility of Kitw/Kitwv mouse with nonobstructive azoospermia through gene editing on SSCs mediated by CRISPR-Cas9

Background Male infertility is a serious social problem in modern society. Nonobstructive azoospermia (NOA) caused by germ cell gene defects is an important reason for male infertility, but effective clinical treatment for this disease has not been established. Methods We choose Kit.sup.w/Kit.sup.wv...

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Bibliographic Details
Published in:Stem cell research & therapy 2019-08, Vol.10 (1), p.271-271, Article 271
Main Authors: Li, Xiaoyu, Sun, Tiecheng, Wang, Xiuxia, Tang, Jixin, Liu, Yixun
Format: Article
Language:English
Subjects:
Analysis
Care and treatment
Cloning
CRISPR
Fibroblasts
Gene editing
Gene mutation
Genes
Genetic research
Genome editing
Homology
Infertility
KIT protein
Laboratories
Male infertility
Medical research
Mutation
Plasmids
Point mutation
Reproductive technologies
Science
Spermatogenesis
Stem cell transplantation
Stem cells
Surgery
Zoology
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Summary:Background Male infertility is a serious social problem in modern society. Nonobstructive azoospermia (NOA) caused by germ cell gene defects is an important reason for male infertility, but effective clinical treatment for this disease has not been established. Methods We choose Kit.sup.w/Kit.sup.wv mouse as a research model and try to develop a new treatment strategy and "cure" its infertility. Mutant spermatogonial stem cells (SSCs) were isolated from one single unilateral testis of a 14-day-old Kit.sup.w/Kit.sup.wv mouse and propagated in vitro. The C to T point mutation on Kit.sup.wv site of these SSCs was corrected through CRISPR-Cas9-mediated homology-directed repair (HDR) in vitro. Then, the repaired SSCs were screened out, proliferated, and transplanted into the remaining testis, and complete spermatogenesis was established in the recipient testis. Results Healthy offsprings with wild type Kit gene or Kit.sup.w mutation were obtained through natural mating 4 months after SSC transplantation. Conclusion In this study, we established an effective new treatment strategy for NOA caused by germ cell gene defects through a combination of SSC isolation, CRISPR-Cas9-mediated gene editing, and SSC transplantation, which brought hope for these NOA patients to restore their natural fertility. Keywords: CRISPR-Cas9, Nonobstructive azoospermia, Spermatogonia stem cells
ISSN:1757-6512
1757-6512
DOI:10.1186/s13287-019-1386-7