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

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.

Sperm‐mediated gene transfer: Applications and implications

“…(46,50,51,59,(68)(69)(70) Finally, a novel form of SMGT, termed linker based spermmediated gene transfer (LB-SMGT), has recently been reported. (68) This work employed sperm surface-specific monoclonal antibody (mAb C) complexed with DNA. Capable of specifically binding to the sperm of various species of animals, mAb C served as a linker molecule to attach transgenes to the surface of pig and mice sperm.…”

Effect of the initial dose of foot-and-mouth disease virus on the early viral dynamics within pigs

“…Taking the blood volume to be 1/13 of the pig weight (Chang et al 2002) gives a volume of 1.923 l for a 25 kg pig. The mean cardiac output of a typical pig is 4.5 l min K1 (Åberg et al 2004).…”

Animal Biotechnology III

“…random integration into the host genome, low efficiency and frequent incidence of mosaicism, research has focused on alternate methodologies for improving the generation of transgenic livestock (Kues and Niemann 2004). These include sperm mediated DNA transfer (Gandolfi 1998;Squires 1999;Chang et al 2002;Lavitrano et al 2002), -the intracytoplasmic injection (ICSI) of sperm heads carrying foreign DNA (Perry et al 1999), -injection or infection of oocytes or embryos by viral vectors (Haskell and Bowen 1995;Chan et al 1998;Hofmann et al 2003;Whitelaw et al 2004) or the use of nuclear transfer Cibelli et al 1998;Baguisi et al 1999).…”