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In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses

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Selecting Fully-Modified XNA Aptamers Using Synthetic Genetics.

Alexander I Taylor¹, Philipp Holliger¹

¹Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom.

Current Protocols in Chemical Biology

|June 22, 2018

Summary

This summary is machine-generated.

Synthetic genetics enables the creation of xeno nucleic acid (XNA) aptamers. This method uses cross-chemistry selective exponential enrichment (X-SELEX) to evolve XNA aptamers for specific molecular targets.

Keywords:

SELEX aptamers synthetic genetics xeno nucleic acids (XNA)

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Area of Science:

Synthetic biology
Molecular biology
Biochemistry

Background:

Xeno nucleic acids (XNAs) are artificial DNA/RNA analogs with modified backbones or sugars.
Directed evolution is crucial for developing novel molecular ligands.
XNA aptamers offer potential for diverse molecular recognition applications.

Purpose of the Study:

To describe the application of synthetic genetics for directed evolution of XNA aptamers.
To present the cross-chemistry selective exponential enrichment (X-SELEX) approach.
To detail protocols for XNA repertoire synthesis and functional selection.

Main Methods:

Synthesis of large-sequence XNA libraries (10^14 molecules) using DNA templates.
Selection of functional XNA aptamers via solid-phase immobilization or solution-phase gel shift assays.
Reverse transcription of XNA for subsequent cDNA amplification and sequencing.

Main Results:

Demonstrated a method for generating diverse XNA repertoires.
Successfully isolated functional XNA aptamers specific for target molecules.
Established protocols for XNA synthesis, selection, and analysis.

Conclusions:

The described X-SELEX method is broadly applicable for evolving fully-modified XNA aptamers.
This approach facilitates the selection of XNA aptamers against a wide range of targets.
Synthetic genetics provides a powerful platform for creating novel aptamer-based tools.