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Highly Multiplexed RNA Aptamer Selection using a Microplate-based Microcolumn Device.

Sarah J Reinholt¹, Abdullah Ozer², John T Lis²

¹School of Applied and Engineering Physics, Cornell University, Ithaca, NY 14853, USA.

Scientific Reports

|July 20, 2016

Summary

This summary is machine-generated.

This study introduces MEDUSA, a device for rapid, simultaneous RNA aptamer selection against 19 targets. This method significantly reduces time and reagents, enabling high-affinity aptamer discovery and protein inhibition.

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Primer-Free Aptamer Selection Using A Random DNA Library

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Published on: July 26, 2010

Area of Science:

Biotechnology
Molecular Biology
Biochemistry

Background:

Traditional aptamer selection is time-consuming and reagent-intensive.
Multiplexed selection methods are needed to accelerate aptamer discovery.
High-throughput screening is crucial for identifying functional aptamers.

Purpose of the Study:

To develop a multiplexed RNA aptamer selection method using a microcolumn-based device (MEDUSA).
To reduce the time and reagents required for aptamer selection.
To discover high-affinity aptamers with inhibitory functions against target proteins.

Main Methods:

Utilized MEDUSA, a microplate-based enrichment device, for simultaneous selection of RNA aptamers against 19 targets.
Employed a modified selection process with MEDUSA's reconfigurable microcolumns (parallel and serial connections).
Leveraged 96-well microplate compatibility for high-throughput downstream processing.

Main Results:

Achieved the equivalent of a 10-cycle selection in the time of 4 traditional cycles.
Discovered several aptamers with nanomolar dissociation constants.
Identified aptamers that inhibit mouse decapping exoribonuclease (DXO) activity with low micromolar concentrations.

Conclusions:

MEDUSA and the modified protocol significantly enhance the efficiency of aptamer selection.
This platform facilitates high-throughput aptamer discovery and can be automated.
Developed functional aptamers capable of inhibiting protein activity, demonstrating therapeutic potential.