Safe and efficient ecotropic and amphotropic packaging lines for use in gene transfer experiments.
The construction of a retrovirus packaging cell line which produces high-titer, helper-free retrovirus is an essential prerequisite for experiments whose goal is gene therapy. We have constructed an ecotropic packaging cell line, GP + E-86, and an amphotropic packaging cell line, GP + envAm12, in which the viral gag and pol genes are on one plasmid and the viral env gene is on another plasmid. Both plasmids contain deletions of the psi packaging sequence and the 3' LTR. The fragmented helper virus genomes, when introduced into 3T3 cells, produce titers of delta neo or N2 retrovirus which are comparable to titers produced from packaging cells containing the helper virus genome on a single plasmid. We have found no evidence for the generation of wild-type retrovirus using the GP + E-86 and GP + envAm12 packaging lines, either alone or in combination with the N2 retroviral vector. These results indicate that these packaging lines are safe and efficient for use in experiments designed for murine (using GP + E-86) and human (using GP + envAm12) gene therapy.
Markowitz
D G
DG
Department of Genetics and Development, Columbia University, College of Physicians and Surgeons, New York, New York 10032.
Goff
S P
SP
Bank
A
A
D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant; D004279:DNA, Viral; D015752:Genes, env; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D010957:Plasmids; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection
D004274:DNA, Recombinant; D004279:DNA, Viral