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Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA.

Publication ,  Journal Article
Wicks, SL; Hargrove, AE
Published in: Methods (San Diego, Calif.)
September 2019

Fluorescent indicator displacement (FID) assays are an advantageous approach to convert receptors into optical sensors that can detect binding of various ligands. In particular, the identification of ligands that bind to RNA receptors has become of increasing interest as the roles of RNA in cellular processes and disease pathogenesis continue to be discovered. Small molecules have been validated as tools to elucidate unknown RNA functions, underscoring the critical need to rapidly identify and quantitatively characterize RNA:small molecule interactions for the development of chemical probes. The successful application of FID assays to evaluate interactions between diverse RNA receptors and small molecules has been facilitated by the characterization of distinct fluorescent indicators that reversibly bind RNA and modulate the fluorescence signal. The utility of RNA-based FID assays to both academia and industry has been demonstrated through numerous uses in high-throughput screening efforts, structure-activity relationship studies, and in vitro target engagement studies. Furthermore, the development, optimization, and validation of a variety of RNA-based FID assays has led to general guidelines that can be utilized for facile implementation of the method with new or underexplored RNA receptors. Altogether, the use of RNA-based FID assays as a general analysis tool has provided valuable insights into small molecule affinity and selectivity, furthering the fundamental understanding of RNA:small molecule recognition. In this review, we will summarize efforts to employ FID assays using RNA receptors and describe the significant contributions of the method towards the development of chemical probes to reveal unknown RNA functions.

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Published In

Methods (San Diego, Calif.)

DOI

EISSN

1095-9130

ISSN

1046-2023

Publication Date

September 2019

Volume

167

Start / End Page

3 / 14

Related Subject Headings

  • Structure-Activity Relationship
  • Small Molecule Libraries
  • RNA
  • Ligands
  • Humans
  • High-Throughput Screening Assays
  • Fluorescent Dyes
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences
 

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Wicks, S. L., & Hargrove, A. E. (2019). Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA. Methods (San Diego, Calif.), 167, 3–14. https://doi.org/10.1016/j.ymeth.2019.04.018
Wicks, Sarah L., and Amanda E. Hargrove. “Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA.Methods (San Diego, Calif.) 167 (September 2019): 3–14. https://doi.org/10.1016/j.ymeth.2019.04.018.
Wicks SL, Hargrove AE. Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA. Methods (San Diego, Calif). 2019 Sep;167:3–14.
Wicks, Sarah L., and Amanda E. Hargrove. “Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA.Methods (San Diego, Calif.), vol. 167, Sept. 2019, pp. 3–14. Epmc, doi:10.1016/j.ymeth.2019.04.018.
Wicks SL, Hargrove AE. Fluorescent indicator displacement assays to identify and characterize small molecule interactions with RNA. Methods (San Diego, Calif). 2019 Sep;167:3–14.
Journal cover image

Published In

Methods (San Diego, Calif.)

DOI

EISSN

1095-9130

ISSN

1046-2023

Publication Date

September 2019

Volume

167

Start / End Page

3 / 14

Related Subject Headings

  • Structure-Activity Relationship
  • Small Molecule Libraries
  • RNA
  • Ligands
  • Humans
  • High-Throughput Screening Assays
  • Fluorescent Dyes
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences