Pig transgenesis by Sleeping Beauty DNA transposition

Area of Science:

• Biomedical Research
• Genetics
• Animal Models

Background:

• Genetically engineered pigs offer valuable models for human disease research and intervention studies.
• Efficient gene insertion methods using non-viral carriers are crucial for developing novel disease models.

Purpose of the Study:

• To create transgenic Göttingen minipigs using Sleeping Beauty DNA transposition and handmade cloning.
• To assess the efficiency and characteristics of Sleeping Beauty transposition in porcine fibroblasts.

Main Methods:

• Generation of transgenic pigs via Sleeping Beauty DNA transposition in primary porcine fibroblasts.
• Somatic cell nuclear transfer using handmade cloning technique.
• Utilized DNA transposon vectors with a human ubiquitin C promoter for transgene expression.

Main Results:

• Successfully produced transgenic Göttingen minipigs expressing green fluorescent protein.
• Animals exhibited systemic transgene expression with multiple copies (8-13) of the transgene.
• Observed both transposase-catalyzed insertions and random plasmid DNA insertions.

Conclusions:

• Sleeping Beauty DNA transposition can be used to create transgenic pigs for disease modeling.
• Identified challenges including coincidental plasmid insertion and lower transposition activity in porcine cells.
• This technology provides a foundation for future development of porcine disease models.