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Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer. |
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| Authors: | 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 Jr 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 |
| Affiliation: | 2. BioAgri Corporation-Taiwan Branch, Fl. 8-8, No. 8, Song-Chiang Rd., Taipei, Taiwan 3. Department of Chemistry, Soochow University, Taipei, Taiwan 1. BioAgri Corporation, 17711 Rowland St., City of Industry, CA, 91748, U.S.A 4. Department of Anesthesiology, UCLA, CA, 90095, U.S.A 5. Center for Craniofacial Molecular Biology, University of Southern California, CA, 90033, U.S.A 6. Taiwan Livestock Research Institute, Hsinhua, Taiwan 7. Analytical Cytometry Core, Division of Biology, City of Hope, Duarte, CA, 91010, U.S.A 8. Department of Pathology, Tri-Service General Hospital, Neihu, Taipei, Taiwan 9. Institute of Biopharmaceutical Science, National Yang-Ming University, Taipei, Taiwan |
| Abstract: | BackgroundTransgenic 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.ResultsThe 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).ConclusionsOur data suggests that LB-SMGT could be used to generate transgenic animals efficiently in many different species. |
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