Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges.

摘要: Abstract We have constructed a set of packaging cell lines useful for the generation helper-free recombinant retroviruses with amphotropic and ecotropic host ranges. To eliminate problems transfer functions helper virus formation encountered previously available systems, two mutant Moloney murine leukemia virus-derived proviral genomes carrying complementary mutations in gag-pol or env regions were sequentially introduced into NIH 3T3 cells by cotransformation. Both contained deletion psi sequence necessary efficient encapsidation retroviral particles additional alterations at 3' end provirus. We show that resulting CRIP CRE can be used to isolate clones stably produce high titers (10(6) colony-forming units/ml) ranges, respectively. More importantly, we demonstrate viral producers derived from do not functions, yield virus, even under conditions where existing shown and/or virus. These properties make them particularly valuable reagents vivo gene studies aimed lineage analysis development human replacement therapies.