Evaluation of siRNA Stability and Interaction with Serum Components Using an Agarose Gel-Based Single-Molecule FRET Labeling Method

Methods Mol Biol. 2021:2282:43-56. doi: 10.1007/978-1-0716-1298-9_4.

Abstract

Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

Keywords: Agarose gel electrophoresis; Förster resonance energy transfer; RNA interference; Serum protein corona; Small interfering RNA; siRNA stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Proteins / metabolism*
  • Electrophoresis, Agar Gel*
  • Fluorescence Resonance Energy Transfer*
  • Kinetics
  • Luminescent Measurements
  • RNA Interference*
  • RNA Stability*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism*
  • Research Design
  • Single Molecule Imaging*
  • Workflow

Substances

  • Blood Proteins
  • RNA, Small Interfering