Abstract
Genetically encodable sensors have been widely used in the detection of intracellular molecules ranging from metal ions and metabolites to nucleic acids and proteins. These biosensors are capable of monitoring in real-time the cellular levels, locations, and cell-to-cell variations of the target compounds in living systems. Traditionally, the majority of these sensors have been developed based on fluorescent proteins. As an exciting alternative, genetically encoded RNA-based molecular sensors (GERMS) have emerged over the past few years for the intracellular imaging and detection of various biological targets. In view of their ability for the general detection of a wide range of target analytes, and the modular and simple design principle, GERMS are becoming a popular choice for intracellular analysis. In this review, we summarize different design principles of GERMS based on various RNA recognition modules, transducer modules, and reporting systems. Some recent advances in the application of GERMS for intracellular imaging are also discussed. With further improvement in biostability, sensitivity, and robustness, GERMS can potentially be widely used in cell biology and biotechnology.
Highlights
The detection and quantification of cellular proteins, nucleic acids, and metabolites is critical in understanding cellular signaling pathways and many other physiological processes
Capture-Systematic Evolution of Ligands by EXponential enrichment (SELEX) is different from conventional SELEX in that it does not require the immobilization of the target compounds to beads or surfaces [107]
Similar to riboswitches, the identified aptamers in the Capture-SELEX have been already optimized to respond to target binding by changing the RNA conformation, which is important for sensor development
Summary
The detection and quantification of cellular proteins, nucleic acids, and metabolites is critical in understanding cellular signaling pathways and many other physiological processes. An alternative class of RNA-based fluorescent biosensors has been developed for intracellular applications [29,30,31,32] These Genetically Encoded RNA-based Molecular Sensors (GERMS) consist of three components: a recognition module, a reporting system, and a transducer module. GERMS can be and rationally modified for the detection of a wide range of target molecules with good selectivity and sensitivity These genetically encodable sensors have shown promising potential in detecting intracellular RNAs, proteins, metabolites, signaling molecules, and metal ions [29,30,32,38,39,40,41]. Recent examples will be further provided to demonstrate the intracellular applications of these novel RNA-based sensors
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