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In vitro Selection of Chemically Modified DNAzymes
Po-Jung Jimmy Huang1, Juewen Liu1
1Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario, N2L 3G1, Canada.
Chemistryopen
|October 26, 2020
Summary
Modified DNAzymes (catalytic DNA) with enhanced chemical functions were developed using in vitro selection and chemical modifications. These novel DNAzymes show specific RNA cleavage activity, metal ion activation, and applications in biosensing.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- DNAzymes are DNA molecules with catalytic functions, primarily studied for RNA cleavage.
- Chemical modifications expand DNAzyme capabilities beyond natural DNA.
- In vitro selection is a key technique for discovering and engineering DNAzymes.
Purpose of the Study:
- To review RNA-cleaving DNAzymes modified during in vitro selection.
- To highlight methods for incorporating modified nucleotides into DNAzymes.
- To explore applications of modified DNAzymes in biosensing and molecular probes.
Main Methods:
- In vitro selection incorporating modified nucleotides via PCR or primer extension.
- Introduction of specific modifications like phosphorothioate and histidine-glycine.
- Labeling with fluorophore/quencher pairs for signaling applications.
- Investigation of cleavage for modified RNA substrates (2'-5' and L-RNA).
Main Results:
- DNAzymes activated by specific metal ions (Zn2+, Hg2+, Cu2+, Cd2+, Ni2+) were developed.
- Modified DNAzymes capable of RNA cleavage without divalent metal ions were created.
- Signaling DNAzymes for detecting metal ions and cells were successfully engineered.
- Cleavage of non-natural RNA linkages (2'-5' and L-RNA) by modified DNAzymes was demonstrated.
Conclusions:
- Chemical modifications significantly enhance the functionality and specificity of RNA-cleaving DNAzymes.
- Modified DNAzymes offer versatile platforms for biosensing, diagnostics, and biochemical research.
- Further research into modified DNAzymes holds promise for advanced molecular tools and therapeutic applications.
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