Figure 1

Diagrams of the retroviral vectors and the shuttle-packaging scheme. A) The MMP-based vector series contains GFP transcriptionally regulated by the MPSV LTR and the control or 3 different HIV-1 specific RT aptamers transcriptionally regulated by the U6+1 promoter positioned in the antisense orientation. The forward and reverse arrows above GFP and the 3' UTR represent the PCR primers used to amplify the inhibitor cassette in transduced cells. B) Generation of high-titer producer cell lines by shuttle packaging. Vector plasmids were transiently transfected into Phoenix (GaLV) packaging cell line and supernatant was used to transduce Phoenix (amphotropic) packaging cell lines. After selecting for Phoenix (amphotropic) cells expressing GFP, supernatants were collected for titering and to transduce Phoenix (GaLV) packaging cell lines with 2 exposures. Supernatants from these Phoenix (GaLV) cells were collected for titration and to transduce the CD4⁺ cell line CEMx174.