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A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay
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A massively parallel screening platform for converting aptamers into molecular switches.
Alex M Yoshikawa1, Alexandra E Rangel2, Liwei Zheng2
1Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA.
Nature Communications
|April 24, 2023
Summary
We developed a new screening method to create aptamer molecular switches without needing to know their structure. This approach generates sensitive, fast-acting switches for detecting molecules like glucose.
Area of Science:
- Biotechnology
- Molecular Biology
- Chemical Biology
Background:
- Aptamers are valuable molecular tools that change shape when binding targets.
- Conventional methods struggle to create aptamers with built-in switching capabilities.
- Existing software limitations hinder the rational design of aptamer switches.
Purpose of the Study:
- To develop a generalizable strategy for converting aptamers into molecular switches.
- To bypass the need for structural information or rational design in switch engineering.
- To create novel aptamer switches with high sensitivity and rapid kinetics.
Main Methods:
- A massively parallel screening-based strategy was employed.
- The method enables conversion of aptamers into molecular switches without prior structural knowledge.
- Applied to ATP aptamer and a new glucose aptamer.
Main Results:
- Successfully generated multiple aptamer switches from existing and newly selected aptamers.
- Achieved signal-on switching for ATP and signal-off switching for glucose.
- Demonstrated second-scale kinetics for target binding and switching.
- Developed a glucose-responsive switch with ~30-fold improved sensitivity over previous designs.
Conclusions:
- The described screening approach offers a generalizable method for aptamer switch development.
- This strategy overcomes limitations of in silico modeling and rational design.
- Enables creation of diverse aptamer switches for various applications including biosensing and diagnostics.
