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Published on: November 5, 2012
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Cas9-expressing cattle using the PiggyBac transposon all-in-one system.
Dong-Hyeok Kwon1,2, Gyeong-Min Gim1,3, Soo-Young Yum1,3
1Department of Theriogenology, College of Veterinary Medicine and the Research Institute for Veterinary Science, Seoul National University, Seoul, 08826, Republic of Korea.
BMC Genomics
|March 5, 2025
Summary
Researchers successfully created CRISPR/Cas9-expressing cattle, enabling efficient gene editing for improved bovine genetics and disease resistance. This breakthrough advances cattle genetic engineering and disease modeling, paving the way for new biotechnological applications.
Area of Science:
- Biotechnology
- Genomics
- Animal Science
Background:
- Genetic engineering in cattle is challenging due to long gestation, single pregnancies, and high costs.
- Slow germline transmission validation and lack of germline-competent embryonic stem cells impede progress.
- Recent advances in genome editing technologies (CRISPR-Cas9) have enabled successful production of Cas9-expressing animals in other species.
Purpose of the Study:
- To generate CRISPR/Cas9-expressing cattle as a resource for bovine genome editing.
- To advance the understanding of bovine genetics and disease resistance.
- To establish novel transgenic bovine models for research.
Main Methods:
- Production of two types of Cas9-expressing cattle: Cas9-RFP-Fatty acid dehydrogenase I (FatI) and Cas9-GFP-sgRNA for the prion protein (sgPRNP).
- Transfection of somatic cells with single-guide RNAs (sgRNAs) to induce target gene mutations.
- Cryopreservation of semen from Cas9-expressing males for fertilization of wild-type oocytes.
- In vitro production and gene editing evaluation in embryos derived from F1 semen.
Main Results:
- Successfully produced Cas9-expressing cattle (F0 generation) and confirmed transgene transmission to the next generation (F1 calves and F2 embryos).
- Demonstrated gene editing capabilities (knockout and knock-in) in somatic cells and embryos using CRISPR/Cas9.
- Established PRNP-mutated F1 cattle as a potential model for bovine spongiform encephalopathy resistance.
Conclusions:
- Successfully generated the first Cas9-expressing cattle with heritable transgenes.
- Validated the potential for in vitro and in vivo genome editing using somatic and germ cells from multiple generations.
- These transgenic cattle models offer a valuable resource for advancing bovine genomics, disease resistance research, and biotechnology.
