Bacteria contain a diverse set of RNAs to provide tight regulation of gene expression in response to environmental stimuli. Bacterial small RNAs (sRNAs) work in conjunction with protein cofactors to bind complementary mRNA sequences in the cell, leading to up‐ or downregulation of protein synthesis. In vivo imaging of sRNAs can aid in understanding their spatiotemporal dynamics in real time, which inspires new ways to manipulate these systems for a variety of applications including synthetic biology and therapeutics. Current methods for sRNA imaging are quite limited in vivo and do not provide real‐time information about fluctuations in sRNA levels. Herein, we describe our efforts toward the development of an RNA‐based fluorescent biosensor for bacterial sRNA both in vitro and in vivo. We validated these sensors for three different bacterial sRNAs in Escherichia coli and demonstrated that the designs provide a bright, sequence‐specific signal output in response to exogenous and endogenous RNA targets.

中文翻译：

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用于细菌 sRNA 活细胞检测的 RNA 荧光生物传感器

细菌含有多种 RNA，可根据环境刺激对基因表达进行严格调控。细菌小 RNA (sRNA) 与蛋白质辅助因子结合，结合细胞中的互补 mRNA 序列，从而上调或下调蛋白质合成。sRNA 的体内成像有助于实时了解其时空动态，从而激发了操纵这些系统用于合成生物学和治疗学等各种应用的新方法。目前的 sRNA 成像方法在体内相当有限，并且不能提供有关 sRNA 水平波动的实时信息。在此，我们描述了我们在体外和体内开发基于 RNA 的细菌 sRNA 荧光生物传感器的努力。我们针对大肠杆菌中的三种不同细菌 sRNA 验证了这些传感器，并证明这些设计可提供明亮的序列特异性信号输出来响应外源和内源 RNA 靶标。