Efficient generation of bone morphogenetic protein 15-edited Yorkshire pigs using CRISPR/Cas9†

Journal Article · Thu Aug 06 00:00:00 EDT 2020 · Biology of Reproduction

DOI:https://doi.org/10.1093/biolre/ioaa138· OSTI ID:1646541

Shi, Xuan ^[1]; Tang, Tao ^[1]; Lin, Qiyuan ^[1]; Liu, Hongbo ^[1]; Qin, Yufeng ^[1]; Liang, Xinyu ^[1]; Cong, Peiqing ^[1]; Mo, Delin ^[1]; Liu, Xiaohong ^[1]; Chen, Yaosheng ^[1]; He, Zuyong ^[1]

---

1. State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, P. R. China

Abstract

Bone morphogenetic protein 15 (BMP15), a member of the transforming growth factor beta (TGF-β) superfamily, plays an essential role in ovarian follicular development in mono-ovulatory mammalian species. Studies using a biallelic knockout mouse model revealed BMP15 potentially has just a minimal impact on female fertility and ovarian follicular development in poly-ovulatory species. In contrast, our previous study demonstrated that in vivo knockdown of BMP15 significantly affected porcine female fertility, as evidenced by the dysplastic ovaries containing significantly decreased numbers of follicles and an increased number of abnormal follicles. This finding implied that BMP15 plays an important role in the regulation of female fertility and ovarian follicular development in poly-ovulatory species. To further investigate the regulatory role of BMP15 in porcine ovarian and follicular development, here, we describe the efficient generation of BMP15-edited Yorkshire pigs using CRISPR/Cas9. Using artificial insemination experiments, we found that the biallelically edited gilts were all infertile, regardless of different genotypes. One monoallelically edited gilt #4 (Δ66 bp/WT) was fertile and could deliver offspring with a litter size comparable to that of wild-type gilts. Further analysis established that the infertility of biallelically edited gilts was caused by the arrest of follicular development at pre-antral stages, with formation of numerus structurally abnormal follicles, resulting in streaky ovaries and the absence of obvious estrous cycles. Our results strongly suggest that the role of BMP15 in non-rodent poly-ovulatory species may be as important as that in mono-ovulatory species.

• Sponsoring Organization: USDOE
• OSTI ID: 1646541
• Journal Information: Biology of Reproduction, Journal Name: Biology of Reproduction; ISSN 0006-3363
• Publisher: Oxford University Press
• Country of Publication: United States
• Language: English