期刊
出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.2022562118
关键词
Cas9 transgenic chicken; Cas9 transgenic pig; in vivo genome editing; CRISPR-Cas9
资金
- Deutsche Forschungsgemeinschaft (DFG) [Schu2446/4-1, KA3492/6-1, F L948/1-1]
- DFG Collaborative Research Center [Transregio 267]
- DFG Collaborative Research Center 1321
- German Center for Cardiovascular Research on Gene therapy for neonatal sarcomeric cardiomyopathies: Towards first-in -patient [81X2600413]
Research on genetically modified animals has mostly focused on mice, but also includes species like pigs that are more physiologically similar to humans, and cross-species comparisons with phylogenetically distant species like chickens. CRISPR-Cas9 is a versatile genetic editing method applicable across various species. The successful generation of transgenic chickens and pigs that express Cas9 in all organs has confirmed the functionality of Cas9 across different target genes, cell types, and in vivo applications.
Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.
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