Versatile generation of precise gene edits in bovines using SEGCPN

Area of Science:

• Animal Biotechnology
• Genomics
• Gene Editing

Background:

• Genome editing systems are established for large farm animals.
• Precise gene editing techniques like targeted deletion, gene tagging, and large fragment replacement are challenging in these animals.

Purpose of the Study:

• To develop a versatile gene-editing technology for precise modifications in large farm animals.
• To demonstrate the efficacy of the new technology for various gene editing applications in bovine.

Main Methods:

• Established versatile self-excising gene-targeting technology in combination with programmable nucleases (SEGCPN).
• Applied SEGCPN to generate bovine embryos with point mutations and deletions at the MSTN locus.
• Utilized SEGCPN for EGFP labeling at the SRY locus in bulls and large gene replacement in cows.

Main Results:

• Successfully generated bovine embryos with precise mutations at the MSTN locus.
• Achieved successful EGFP gene tagging in the SRY locus of bulls.
• Generated humanized cows by replacing the endogenous α-casein gene with human α-lactalbumin.

Conclusions:

• The SEGCPN method provides a versatile platform for diverse precise gene editing in large animals.
• This technology has significant potential applications in agriculture and the development of disease models.