PGC-based cryobanking, regeneration through germline chimera mating, and CRISPR/Cas9-mediated TYRP1 modification in indigenous Chinese chickens
Primordial germ cells (PGCs) are vital for producing sperm and eggs and are crucial for conserving chicken germplasm and creating genetically modified chickens. However, efforts to use PGCs for preserving native chicken germplasm and genetic modification via CRISPR/Cas9 are limited. Here we show that we established 289 PGC lines from eight Chinese chicken populations with an 81.6% success rate. We regenerated Piao chickens by repropagating cryopreserved PGCs and transplanting them into recipient chickens, achieving a 12.7% efficiency rate. These regenerated chickens carried mitochondrial DNA from female donor PGC and the rumplessness mutation from both male and female donors. Additionally, we created the TYRP1 (tyrosinase-related protein 1) knockout (KO) PGC lines via CRISPR/Cas9. Transplanting KO cells into male recipients and mating them with wild-type hens produced four TYRP1 KO chickens with brown plumage due to reduced eumelanin production. Our work demonstrates efficient PGC culture, cryopreservation, regeneration, and gene editing in chickens.
Authors
- Yuya Okuzaki (ORCID: https://orcid.org/0000-0002-7212-4366)
- Feizhou Sun (ORCID: https://orcid.org/0009-0007-8589-1023)
- Shusei Mizushima (ORCID: https://orcid.org/0000-0002-1176-1994)
- Yoshiaki Nakamura (ORCID: https://orcid.org/0000-0002-5387-1630)
- Takayuki Suzuki (ORCID: https://orcid.org/0000-0003-2364-0935)
- Kaixiang Xu (ORCID: https://orcid.org/0000-0001-6665-3360)
- Fangxian Zhu
- Kumiko Tanabe (ORCID: https://orcid.org/0000-0002-3580-5701)
- Hong‐Jiang Wei (ORCID: https://orcid.org/0000-0002-5663-1093)
- Yanhua Su (ORCID: https://orcid.org/0000-0002-2265-3390)
- Taiyun Wei
- Muhammad Ameen Jamal
- Gang Liu (ORCID: https://orcid.org/0000-0001-6313-7758)
- Samiullah Khan (ORCID: https://orcid.org/0000-0002-0948-5003)
- Hong‐Ye Zhao (ORCID: https://orcid.org/0000-0002-5900-3033)
- Mingshan Wang (ORCID: https://orcid.org/0000-0003-4663-2735)
- Keiji Kinoshita
- Heng Zhao (ORCID: https://orcid.org/0000-0002-9320-5894)
- Ken‐ichi Nishijima (ORCID: https://orcid.org/0000-0002-7189-465X)
Institutions
- Hiroshima University (JP)
- Kunming Institute of Zoology (CN)
- Yunnan University (CN)
- Yunnan Agricultural University (CN)
- Chinese Academy of Sciences (CN)
- Hokkaido University (JP)
- National Animal Husbandry Service (CN)
- Osaka Metropolitan University (JP)
- Nagoya University (JP)
- Tokyo Metropolitan University (JP)
Publication Details
- Journal
- Communications Biology
- Published
- 2024-09-13
- DOI
- https://doi.org/10.1038/s42003-024-06775-5
- Citations
- 10
- Primary Topic
- Animal Genetics and Reproduction
- Type
- article
- Field-Weighted Citation Impact
- 5.50
Funders
- Major Science and Technology Projects in Yunnan Province