Optimization of RNA-based c-di-GMP fluorescent sensors through tuning their structural modules

“…Such genetically-encoded biosensors were developed, for instance, to detect SAM [116], FMN [117], TPP, cyclic AMP [118], as well as various types of cyclic dinucleotides [119][120][121][122][123][124][125]. In general, these sensors are based on the transient destabilization strategy (Figure 5h) similar to that described above for RNA and protein detection.…”

Section: Detection Of Other Biological Molecules and Ionsmentioning

confidence: 99%

Development and Applications of Fluorogen/Light-Up RNA Aptamer Pairs for RNA Detection and More

Ryckelynck

2020

Methods in Molecular Biology

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The central role of RNA in living systems made it highly desirable to have non-invasive and sensitive technologies allowing for imaging the synthesis and the location of these molecules in living cells. This need motivated the development of small pro-e ce ec e ca ed e a bec e e ce b d e e ca e c dab e RNA ca ed-a a e. Ye , e development of these fluorogen/light-up RNA pairs is a long and thorough process starting with the careful design of the fluorogen and pursued by the selection of a specific and efficient synthetic aptamer. This chapter summarizes the main design and the selection strategies used up to now prior to introducing the main pairs. Then, the vast application potential of these molecules for live-cell RNA imaging and other applications will be presented and discussed.

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“…This strategy was originally pioneered using ribozymes as reporting modules [ 101 ] and early work with MG aptamer allowed establishing the proof-of-concept experiment using light-up aptamers as reporter [ 84 ]. Since then, Spinach RNA-based fluorogenic biosensors have been developed to specifically report on the presence of metabolites such as FMN [ 84 ], cyclic-di-GMP [ 85 , 102 , 103 , 104 ], cyclic-di-AMP [ 105 ], cyclic AMP-GMP [ 106 , 107 ], cyclic-AMP [ 108 ], S-adenosylmethionine (SAM) [ 109 ], S-adenosyl- l -homocysteine (SAH) [ 110 ], Thiamine Pyrophosphate (TPP) [ 111 ] and neurotransmitter precursors [ 112 ]. The majority of these biosensors are built using riboswitch-derived aptamers.…”

Section: Applications Of Rna-based Fluorogenic Modulesmentioning

confidence: 99%

Light-Up RNA Aptamers and Their Cognate Fluorogens: From Their Development to Their Applications

Bouhedda

Autour

Ryckelynck

2017

IJMS

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An RNA-based fluorogenic module consists of a light-up RNA aptamer able to specifically interact with a fluorogen to form a fluorescent complex. Over the past decade, significant efforts have been devoted to the development of such modules, which now cover the whole visible spectrum, as well as to their engineering to serve in a wide range of applications. In this review, we summarize the different strategies used to develop each partner (the fluorogen and the light-up RNA aptamer) prior to giving an overview of their applications that range from live-cell RNA imaging to the set-up of high-throughput drug screening pipelines. We then conclude with a critical discussion on the current limitations of these modules and how combining in vitro selection with screening approaches may help develop even better molecules.

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Optimization of RNA-based c-di-GMP fluorescent sensors through tuning their structural modules

Cited by 4 publications

References 34 publications

A STING-based fluorescent polarization assay for monitoring activities of cyclic dinucleotide metabolizing enzymes

A STING-based fluorescent polarization assay for monitoring activities of cyclic dinucleotide metabolizing enzymes

Development and Applications of Fluorogen/Light-Up RNA Aptamer Pairs for RNA Detection and More

Light-Up RNA Aptamers and Their Cognate Fluorogens: From Their Development to Their Applications

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