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

Keejong Chang^*²^,3 , Jin Qian^*¹ , MeiSheng Jiang^*⁴ , Yi-Hsin Liu^*⁵ , Ming-Che Wu⁶ , Chi-Dar Chen² , Chao-Kuen Lai² , Hsin-Lung Lo³ , Chin-Ton Hsiao² , Lucy Brown⁷ , James Bolen Jr⁷ , Hsiao-I Huang¹ , Pei-Yu Ho² , Ping Yao Shih² , Chen-Wen Yao⁸ , Wey-Jinq Lin⁹ , Chung-Hsi Chen³ , Fang-Yi Wu³ , Yi-Jen Lin³ , Jing Xu¹ and Ken Wang¹

¹BioAgri Corporation, 17711 Rowland St., City of Industry, CA 91748, U.S.A

²BioAgri Corporation-Taiwan Branch, Fl. 8-8, No. 8, Song-Chiang Rd., Taipei, Taiwan

³Department of Chemistry, Soochow University, Taipei, Taiwan

⁴Department of Anesthesiology, UCLA, CA 90095, U.S.A

⁵Center for Craniofacial Molecular Biology, University of Southern California, CA 90033, U.S.A

⁶Taiwan Livestock Research Institute, Hsinhua, Taiwan

⁷Analytical Cytometry Core, Division of Biology, City of Hope, Duarte, CA 91010, U.S.A

⁸Department of Pathology, Tri-Service General Hospital, Neihu, Taipei, Taiwan

⁹Institute of Biopharmaceutical Science, National Yang-Ming University, Taipei, Taiwan

author email corresponding author email* Contributed equally

BMC Biotechnology 2002, 2:5doi:10.1186/1472-6750-2-5


Published:	19 April 2002

Abstract

Background

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.

Results

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).

Conclusions

Our data suggests that LB-SMGT could be used to generate transgenic animals efficiently in many different species.