Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer.
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.
Chang
Keejong
K
BioAgri Corporation-Taiwan Branch, Fl. 8-8, No. 8, Song-Chiang Rd., Taipei, Taiwan. kchang@bioagricorp.com.tw
Qian
Jin
J
Jiang
MeiSheng
M
Liu
Yi-Hsin
YH
Wu
Ming-Che
MC
Chen
Chi-Dar
CD
Lai
Chao-Kuen
CK
Lo
Hsin-Lung
HL
Hsiao
Chin-Ton
CT
Brown
Lucy
L
Bolen
James
J
Huang
Hsiao-I
HI
Ho
Pei-Yu
PY
Shih
Ping Yao
PY
Yao
Chen-Wen
CW
Lin
Wey-Jinq
WJ
Chen
Chung-Hsi
CH
Wu
Fang-Yi
FY
Lin
Yi-Jen
YJ
Xu
Jing
J
Wang
Ken
K
D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002417:Cattle; D002645:Chickens; D005260:Female; D005307:Fertilization in Vitro; D018014:Gene Transfer Techniques; D006041:Goats; D006801:Humans; D007315:Insemination, Artificial / Q000379:methods; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010057:Oviducts / Q000601:surgery; D012756:Sheep; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism*; D013552:Swine
D000911:Antibodies, Monoclonal