pR8.71 HIV-1减毒载体R8.71质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

pR8.71 HIV-1减毒载体R8.71质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

As a best approximation of an RCL, researchers used a virus derived from HIV, R8.71, multiply attenuated by inactivation of the four accessory genes vif, vpr, vpu, and nef, and phenotypically pseudotyped by VSV G, allowing its first round of infection to proceed through the VSV envelope and the subsequent infections to occur through the HIV envelope. This phenotypic pseudotyping allows one to approximate a potential RCL better while avoiding the generation of an HIV strain genetically pseudotyped with VSV G and the biosafety concerns that such a virus would pose. We have used this phenotypically pseudotyped virus as an RCL standard to develop an RCL detection assay with a sensitivity and volume adapted for the testing of large-scale productions of HIV-based lentiviral vector.
The positive control virus stock, R8.71 transiently VSV G pseudotyped, was prepared by cotransfecting 20 μg of the pR8.71 plasmid and 15 μg of the pMD.G envelope plasmid. Among the combinations tested, a 20:15 ratio of pR8.71 to pMD.G yielded virus with the highest infectivity.


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