Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer.

Keejong Chang, Jin Qian, MeiSheng Jiang, Yi-Hsin Liu, Ming-Che Wu, Chi-Dar Chen, Chao-Kuen Lai, Hsin-Lung Lo, Chin-Ton Hsiao, Lucy Brown, James Bolen, Hsiao-I Huang, Pei-Yu Ho, Ping Yao Shih, Chen-Wen Yao, Wey-Jinq Lin, Chung-Hsi Chen, Fang-Yi Wu, Yi-Jen Lin, Jing Xu, Ken Wang

Transgenic animals have become valuable tools for both research and applied purposes. The current method of gene transfer, microinjection, which is widely used in transgenic mouse production, has only had limited success in producing transgenic animals of larger or higher species. Here, we report a linker based sperm-mediated gene transfer method (LB-SMGT) that greatly improves the production efficiency of large transgenic animals. The linker protein, a monoclonal antibody (mAb C), is reactive to a surface antigen on sperm of all tested species including pig, mouse, chicken, cow, goat, sheep, and human. mAb C is a basic protein that binds to DNA through ionic interaction allowing exogenous DNA to be linked specifically to sperm. After fertilization of the egg, the DNA is shown to be successfully integrated into the genome of viable pig and mouse offspring with germ-line transfer to the F1 generation at a highly efficient rate: 37.5% of pigs and 33% of mice. The integration is demonstrated again by FISH analysis and F2 transmission in pigs. Furthermore, expression of the transgene is demonstrated in 61% (35/57) of transgenic pigs (F0 generation). Our data suggests that LB-SMGT could be used to generate transgenic animals efficiently in many different species.