Figure 4

Treatment with Drexel gRNAs individually and together resulted in a significant reduction in LTR-driven transactivation. The HIV reporter cell lines TZM-bl (A) and P4R5 (B) were transfected with Cas9 and individual or packages of gRNAs concurrently with TatIIIB for 48 hr. Cells were then measured for viability (MTT, red bars) and reduction in LTR-driven ß-gal expression. In both cell types, Drexel gRNAs individually or together were able to effectively target and reduce LTR driven activity. C) P4R5 cells were transfected with Cas9 and different gRNAs concurrently with the fully infectious HIV-1 molecular clone pLAI. Cells were then measured for viability and reduction in LTR-driven ß-gal expression. Similarly, the Drexel gRNAs, individually or all together, were able to reduce viral gene expression and LTR driven expression through a number of proposed mechanisms. Statistical significance between Cas9/EV and experimental gRNAs was determined using a one-tailed, one-sample T-test and an *indicated p-values <0.05. Statistical significance between two gRNAs was determined by 2-tailed T-test comparing each item to the Cas9/EV samples. Each dot represents the average of four technical replicates.