Figure 2

Construction of the antiviral miRNA transcripts. (A) The siRNAs A and B were extended and inserted into the miR-30a backbone; siRNA C was extended and inserted into the miR-155 backbone. Bulges that existed in mature wild-type miRNA were eliminated in antiviral miRNAs. Extended sequences in 22 bp siRNAs are in lowercase, whereas native mature miRNA sequence or inserted siRNA sequences are boxed. (B) Screening of effective antiviral miRNAs. The 239FT cells were co-transfected with 500 ng HIV-1 molecular clone pNL4-3, 50 ng pRL, and 100 ng miRNA constructs. The miR-LacZ against β-galactosidase was used as the negative control, whereas shRNAs expressing original 19 bp siRNA was the positive control. CA-p24 levels in the culture supernatant were measured 2 days post-transfection. CA-p24 expression in the presence of miR-LacZ was normalized at 100%. Renilla luciferase was used for normalization of transfection efficiency. Error bars represent the standard deviation of three independent experiments.