Thermoreversible Control of Nucleic Acid Structure and Function with Glyoxal Caging
Steve D Knutson1, Aimee A Sanford1, Colin S Swenson1
1Department of Chemistry, Emory University, Atlanta, Georgia 30322, United States.
Journal of the American Chemical Society
|October 5, 2020
Summary
Glyoxal modification provides reversible heat control for diverse nucleic acids, enabling applications in therapeutics and biocomputing. This method enhances PCR specificity and controls gene expression in living cells.
Area of Science:
- Synthetic Biology
- Biochemistry
- Molecular Biology
Background:
- Controlling nucleic acid structure and activity is crucial for applications in therapeutics, biosensing, nanotechnology, and biocomputing.
- Existing methods for controlling nucleic acids are limited to small-molecule or light stimuli, with heat-triggered control being largely unexplored.
- Current technologies are often restricted to natural nucleic acids and incompatible with polymerase-generated sequences.
Purpose of the Study:
- To develop a method for heat-triggered control of nucleic acid structure and activity.
- To address the limitations of existing responsive nucleic acid technologies.
- To explore the use of glyoxal as a versatile tool for imparting reversible thermal responsiveness to nucleic acids.
Main Methods:
- Glyoxal modification of various DNA and RNA constructs to covalently attach to nucleobases.
- Characterization of glyoxal modification's impact on nucleic acid structure and function.
- Assessment of glyoxal adduct removal kinetics for tunable thermal activity restoration.
- Application of glyoxal caging in RNA aptamers, TNA, PNA scaffolds, CRISPR-Cas9 systems, PCR, and antisense oligonucleotides.
Main Results:
- Glyoxal modification effectively and reversibly modulates the structure and activity of diverse nucleic acid scaffolds.
- Tunable thermal removal of glyoxal adducts allows for controlled restoration of nucleic acid function.
- Glyoxal caging demonstrated versatility in controlling enzyme-nucleic acid interactions, CRISPR-Cas9 activity, PCR specificity, and gene expression via antisense oligonucleotides.
- The method is compatible with natural and synthetic nucleic acids, including polymerase-generated sequences.
Conclusions:
- Glyoxalation is a straightforward, scarless method for imparting reversible thermal responsiveness to virtually any nucleic acid architecture.
- This approach addresses a significant need in synthetic biology for programmable nucleic acid components.
- Glyoxalation offers a versatile new tool for applications in medicine, nanotechnology, and biocomputing.
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