Virofection: a new procedure to achieve stable expression of genes transferred into early embryos.
A new procedure, virofection, designed to stabilize the expression of transfected DNA has been developed. It exploits the capacity of retroviruses to integrate their genome into the chromosomes of host cells. The co-transfection of two plasmids, one carrying the genome of a defective retrovirus vector, the other one encoding all the retroviral proteins, results in a transient production of infectious virus particles. These particles can infect the neighboring cells and this leads to the stable integration of the vector genome. This procedure is time-saving and appears to be quite efficient. When applied to chicken embryonic fibroblasts cultured in vitro, it resulted in the stable expression of the lacZ gene in more than 30% of the cells, and did not induce chronic viremia. Stable lacZ expression was also achieved in chicken embryos in ovo. Virofection appears to be a promising and generally applicable method for implementing stable, safe and efficient gene transfer in vitro and in vivo.
Flamant
F
F
Laboratoire de Biologie Cellulaire et Moléculaire, Ecole Normale Supérieure de Lyon, France.
Demeneix
B
B
Benoist
C
C
Markossian-Belin
S
S
Samarut
J
J
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D010957:Plasmids; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase