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An In Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display
Matthew R Dunn1, John C Chaput
1School of Life Sciences, Arizona State University, Tempe, Arizona; The Biodesign Institute at Arizona State University, Tempe, Arizona.
Current Protocols in Nucleic Acid Chemistry
|June 26, 2014
Summary
Threose nucleic acid (TNA) is an artificial genetic polymer that can pair with DNA and RNA. Researchers developed a DNA display method to evolve functional TNA molecules in the lab.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- Threose nucleic acid (TNA) is a synthetic genetic polymer.
- TNA can form duplex structures and interact with DNA and RNA.
- TNA adopts an A-form helical structure, enabling genetic information exchange.
Purpose of the Study:
- To detail the laboratory evolution of functional TNA molecules.
- To describe the DNA display method for TNA evolution.
- To enable selection of TNA with desired functions.
Main Methods:
- Utilizing an engineered polymerase to copy DNA into TNA.
- Employing the DNA display technique to link TNA phenotype with DNA genotype.
- Physical linkage of TNA molecules to their encoding DNA template.
- Selection and amplification cycles for functional TNA generation.
Main Results:
- Demonstration of a method to evolve functional TNA molecules.
- Successful linkage of TNA function to its encoding DNA sequence.
- Recovery of encoding genetic information via PCR amplification.
- Each selection round takes approximately 3 days.
Conclusions:
- The DNA display method facilitates the laboratory evolution of functional TNA.
- This approach allows for the selection and recovery of TNA aptamers.
- TNA's structural properties support its potential as a genetic material.
