A Fluorogenic RNA-Based Sensor Activated by Metabolite-Induced RNA Dimerization. Academic Article uri icon

Overview

abstract

  • Corn is a fluorogenic RNA aptamer that forms a high-affinity quasi-symmetric homodimer. The Corn dimer interface binds DFHO, resulting in highly photostable yellow fluorescence. Because of its photostability, Corn would be useful in RNA-based small-molecule biosensors, where quantitative accuracy would be affected by photobleaching. Here we describe a strategy for converting the constitutive Corn dimer into a small-molecule-regulated fluorescent biosensor that detects S-adenosylmethionine (SAM) in vitro and in living cells. We fused the Corn aptamer into a helical stem that was engineered by circularly permuting the SAM aptamer from the SAM-III riboswitch. In the absence of SAM, the Corn portion of this fusion RNA is unable to dimerize. However, upon binding SAM, the RNA dimerizes and binds DFHO. This RNA-based biosensor enables detection of SAM dynamics in living mammalian cells. Together, these data describe a class of RNA-based biosensor based on small-molecule-regulated dimerization of Corn.

publication date

  • October 17, 2019

Research

keywords

  • Aptamers, Nucleotide
  • Biosensing Techniques
  • RNA

Identity

PubMed Central ID

  • PMC6939632

Scopus Document Identifier

  • 85076336709

Digital Object Identifier (DOI)

  • 10.1016/j.chembiol.2019.09.013

PubMed ID

  • 31631009

Additional Document Info

volume

  • 26

issue

  • 12