SiRNAs do not induce RNA-dependent transcriptional silencing of retrovirus in human cells

RNA-dependent transcriptional silencing (RdTS) has been reported to operate even in human cell lines. It is tempting to speculate that RdTS plays a role in retroviral gene silencing, considering that retroviral RNA transcripts harbor a U3 promoter sequence that is a potentially good source of double-stranded RNAs. To test this possibility, we constructed several model HeLaS3 cell lines expressing GFP driven by murine leukaemia virus (MLV)-long terminal repeat (LTR) and introduced a series of shRNAs that target the U3 region of the MLV-LTR. However, transcriptional gene silencing was not induced in most instances, in spite of the fact that processed shRNA was found in cellular nuclei, indicating that RdTS does not contribute to MLV gene silencing in host cells.