Conventional MBs have been used to image RNAs in various cellular contexts, but their propensity for nonspecific opening in living cells limits their widespread applications in studies where more sensitive detection is necessary, such as imaging RNA localization and dynamics at the single-molecule level , , . We have recently developed a new MB architecture, known as the 2Me/PSLOOP MB, that elicits a marginal level of FPSs in cells as compared with conventional MBs . We show that 2Me/PSLOOP MBs could accurately image single mRNA transcripts harboring 32 tandem repeats of an MB target sequence using conventional fluorescence microscopy. Currently, RNA dynamics at the single-molecule level has been studied primarily based on engineered RNA molecules that harbor large insertions of MB target or MS2 aptamer sites that potentially interfere with the activities of target RNAs. With further possible approaches for optimizing signal-to-background, such as fluorophore/quencher selection, and the use of more sophisticated imaging techniques, we anticipate that 2Me/PSLOOP MBs can be a promising platform for live-cell, single-molecule imaging of minimally-engineered RNA molecules, or even endogenous RNA molecules, providing researchers with opportunities to study RNAs with unprecedented spatial and temporal resolutions.