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RNA ‐based fluorescent biosensors for live cell detection of bacterial sRNA
Author(s) -
Kitto Rebekah Z.,
Christiansen Kylee E.,
Hammond Ming C.
Publication year - 2021
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.23394
Subject(s) - transfer rna , rna , escherichia coli , synthetic biology , chemistry , in vivo , fluorescence , messenger rna , bacteria , computational biology , microbiology and biotechnology , biology , gene , biochemistry , genetics , physics , quantum mechanics
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.