RNA interference (RNAi), an effective technique for regulating or silencing specific genes, can be applied to treat various diseases. Multiple clinical trials using RNAi are ongoing, and molecular imaging can serve as a powerful tool in RNAi-based therapies. This brief review will highlight the current progress on in vivo imaging of RNAi delivery and silencing effects. Incorporation of suitable molecular imaging techniques into future RNAi-based clinical trials will provide more pieces of the puzzle, thus facilitating the transformation of RNAi into a powerful therapeutic modality in the clinic.