Efficient production of germline transgenic chickens using lentiviral vectors.
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 G2 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.
McGrew
Michael J
MJ
Roslin Institute, Roslin, Midlothian EH25 9PS, UK.
Sherman
Adrian
A
Ellard
Fiona M
FM
Lillico
Simon G
SG
Gilhooley
Hazel J
HJ
Kingsman
Alan J
AJ
Mitrophanous
Kyriacos A
KA
Sang
Helen
H
D000818:Animals; D030801:Animals, Genetically Modified; D015139:Blotting, Southern; D015153:Blotting, Western; D002642:Chick Embryo; D002645:Chickens; D004247:DNA / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D014018:Tissue Distribution; D019076:Transgenes
D004247:DNA