Figure 6.

Effect of six short siRNAs on productive HIV-1 infection including 4 siRNAs targeting HIV-1 promoter region and siRNA HIV-gag, which is known act as PTGS. (A) The sequences within HIV-1 5′LTR targeted by HIV-prom-A-D siRNAs are indicated by highlighting. HIV-prom-B siRNA target sequence includes CpG sites at −159 and −146, HIV-prom-C siRNA target sequence includes CpG at −120, and HIV-prom-D siRNA target sequence includes CpG at −48. CpGs within the 5′LTR of HIV-1 are indicated by emboldened text. Nucleotide numbering is relative to the transcription start site and sequence is that of HIV-1 strain NL4-3. Each siRNA contains at least one CpG site. (B) Effect of siRNAs on the time course of HIV-1 (NL4-3) production in MAGIC-5 cells. At day 7 post-infection siRNAs were transfected into the cells (indicated by arrow). Reverse transcriptase (RT) levels are shown for cultures transfected with HIV-prom-A, B, C, D, HIV-1-gag, and scrambled siRNAs as indicated in the figure legend. (C) HIV-1 proviral DNA is present in cultures transfected with HIV-prom siRNAs regardless of whether infection is productive or not. PCR amplification of proviral DNA extracted from MAGIC-5 cells 38 days after infection (Figure 6B). The amplified region is a 154bp fragment of the HIV-1 gag gene. NC indicates non-infected MAGIC-5 cells.