Abstract
Small RNAs play important roles in regulating gene expression. Here we describe a new approach for characterization and quantification of small regulatory RNAs as well as targeting mRNA at single-cell level by combining single-molecule in situ hybridization and super-resolution imaging. We apply this approach to investigate a stress-induced bacterial small RNA, which is the central regulatory effector of the glucose-phosphate stress response. The quantitative analysis and localization information allow us to establish a kinetic model to describe the sRNA-induced target mRNA degradation in the cell. More importantly, our results demonstrate very promising application of this technique in studying other bacterial small RNA systems, and potentially microRNAs in eukaryotes.
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