Efficient production of germline transgenic chickens using lentiviral vectors

Abstract

An effective method for genetic modification of chickens has yet to be developed. An efficient technology, enabling production of transgenic birds at high frequency and with reliable expression of transgenes, will have many applications, both in basic research and in biotechnology. We investigated the efficiency with which lentiviral vectors could transduce the chicken germ line and examined the expression of introduced reporter transgenes. Ten founder cockerels transmitted the vector to between 4% and 45% of their offspring and stable transmission to the G 2 generation was demonstrated. Analysis of expression of reporter gene constructs in several transgenic lines showed a conserved expression profile between individuals that was maintained after transmission through the germ line. These data demonstrate that lentiviral vectors can be used to generate transgenic lines with an efficiency in the order of 100-fold higher than any previously published method, with no detectable silencing of transgene expression between generations.

Generation of tissue-specific transgenic birds with lentiviral vectors

“…Lentiviral vectors have also been used to generate transgenic pigs and cattle (8,9), and, in principle, they should allow for the generation of transgenic birds. Indeed, promising results were obtained in chickens with the use of recombinant equine infectious anemia virus (EIAV), a type of lentivirus (10). Transgenic chickens were generated that expressed GFP in some tissues; however, the pattern of GFP expression was inconsistent with the expected activity of the viral promoter used.…”

Evaluation of laser-assisted lentiviral transgenesis in bovine

“…Lentiviral vectors are also suitable to create genetically modified birds. Transgenic chicken stably expressed a reporter gene up to the F2 generation (McGrew et al, 2004), and tissue-specific expression in birds was reported in quails, where Scott et al demonstrated that the use of a GFP vector, driven by the human synapsin gene I promoter lead to the expression of GFP in neurons, which was persistent across multiple generations (Scott and Lois, 2005).…”

“…Recent publications have shown that lentiviral vectors are highly efficient for the generation of transgenic mammals (for review see ), such as mouse (Lois et al, 2002a;Pfeifer et al, 2002), rat (Nakagawa et al, 2005), pig (Hofmann, et al 2003 ), cattle (Hofmann et al, 2003;Hofmann et al, 2004) and chicken (McGrew et al, 2004). In our previous studies, we used HIV-1 based lentiviral vectors which contain a self-inactivating (SIN) mutation for the generation of transgenic pigs and cattle (Hofmann et al, 2003a;Hofmann et al, 2004).…”

Relations between animal transgenesis and reproduction

“…These vectors have proven highly efficient in generating transgenic mice [4], pigs [5], cows [6], chickens [7,8] and sheep [9].…”