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A Small-Molecule Approach Enables RNA Aptamers to Function as Sensors for Reactive Inorganic Targets. | LitMetric

AI Article Synopsis

  • Fluorescent light-up aptamer (FLAP) systems are advanced biosensors that can be genetically encoded but typically require in vitro selection for each target detection.
  • This study introduces a small-molecule strategy enabling a single FLAP system to identify unique inorganic and reactive chemicals without needing specific aptamer engineering.
  • The approach utilizes pre-ligands that transform into fluorescent ligands when interacting with reactive inorganics, facilitating whole-cell sensors in live bacteria like E. coli to detect these compounds.

Article Abstract

Fluorescent light-up aptamer (FLAP) systems are promising (bio)sensing platforms that are genetically encodable. However, FLAP-mediated detection of each distinct target necessitates either in vitro selection or engineering of nucleic acid sequences. Furthermore, an aptamer that binds an inorganic target or a chemical species with a short lifetime is challenging to realize. Here, we describe a small-molecule approach that makes it possible for a single FLAP system to detect chemically unique, non-fluorogenic, and reactive inorganics. We developed functionalized pre-ligands of RNA aptamers that bind benzylidene imidazolinones (Baby Spinach, Broccolli, Squash). Reactive inorganics, hydrogen sulfide (HS/HS) and hydrogen peroxide (HO), can specifically convert these pre-ligands into native ligands that fluoresce with FLAPs. Adaptation of this platform to live cells opened an opportunity for constructing whole-cell sensors: Escherichia coli transformed with a Baby Spinach-encoding plasmid and incubated with pre-ligands generated fluorescence in response to exogenous HS/HS or HO. Leveraging the functional group reactivity of small molecules eliminates the requirement of in vitro selection of a new aptamer sequence or oligonucleotide scaffold engineering for distinct molecular targets. Our method allows for detecting inorganic, short-lived species, thereby advancing FLAP systems beyond their current capabilities.

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Source
http://dx.doi.org/10.1002/anie.202421936DOI Listing

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