Native Measurement of Aptamer Folding and Binding Energies Using a Free Energy Shift Assay.

ABSTRACT: Folding and binding energies (ΔG _fold ^° and ΔG _bind ^°) are critical thermodynamic parameters for characterizing the stability and functionality of aptamers. However, it remains difficult to directly characterize aptamer folding and binding in the native environment. Here, we describe a free energy shift assay (FESA) as a simple and instrument-free method to characterize the folding and binding thermodynamics of aptamers. FESA leverages the thermodynamic impact of the aptamer on a panel of toehold-mediated exchange reactions with well-characterized energetics to determine its folding and binding free energies. Using this principle, we successfully determined the ΔG _fold ^° and ΔG _bind

SUBMITTER: Wu Y 

PROVIDER: S-EPMC12308387 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature