Critical Analysis of Rate Constants and Turnover Frequency in Nucleic Acid-Templated Reactions: Reaching Terminal Velocity
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Nucleic acid-templated reactions have attracted significant attention for nucleic acid sensing and imaging. The level of signal amplification obtained from templated reactions is a function of the template turnover, wherein the template acts as the catalyst. Herein, we report the application of a pyridinium linker that immolates upon photocatalytic reduction with a ruthenium complex to yield the fastest nucleic acid templated reaction reported to date. We show that the templated reaction turnover is limited by the duplex dissociation kinetics beyond probes longer than a 6-mer and proceeded fastest for a 5-mer PNA probe. Using a beacon architecture that masks the catalytic template, we show that this methodology can be used for nucleic acid sensing extending the analyte recognition beyond a 5-mer. The system proceeds with a catalytic efficiency of 10 5 M –1 s –1 and achieves turnover frequency of >100 h –1 .
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Templated reactions (ligation or transfer reactions) are possible at much lower concentration, provided the hybridization yields sufficiently long-lived hybridization complex relative to the reaction rate.
“Conversely, templated reactions (ligation or transfer reactions) have been shown to be possible at much lower concentration, provided the hybridization yields sufficiently long-lived hybridization complex relative to the reaction rate.22”
From Sequencing of <scp>d</scp>/<scp>l</scp>-DNA and XNA by Templated-Synthesis (2025) · cites this paper as [22]
The primers used contain chiral PNA residues that hybridize exclusively to d-DNA.
“However, the primers that were used contain chiral PNA residues22,41 that hybridize exclusively to d-DNA.42,43”
From Sequencing of <scp>d</scp>/<scp>l</scp>-DNA and XNA by Templated-Synthesis (2025) · cites this paper as [22]
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Diep, K.-L., Janett, E., Fromm, K. M., & Bochet, C. G. (2025) · The Journal of Organic Chemistry · doi:10.1021/acs.joc.5c00632
One-pot multi-substrate screening of ligation reactions using PNA tags
Kohyama, A., Barluenga, S., & Winssinger, N. (2026) · Chemical Science · doi:10.1039/d5sc08732e
Self-citationMonomer Synthesis and On-Resin Chemical Functionalization of Gamma Peptide Nucleic Acid
Thrikawala, S. W., Aryal, B., & Ly, D. H. (2026) · Methods in molecular biology · doi:10.1007/978-1-0716-5368-5_10
DNA-scaffolded catalysis for aliphatic alcohol oxidation in DEL synthesis
Hartman, E. M., & Martell, J. D. (2026) · Chemical Science · doi:10.1039/d6sc05041g
Toward Ideal Proximity‐Driven Reactions in Chemical Biology
Schauenburg, D. (2026) · ChemBioChem · doi:10.1002/cbic.70565
- Authors
- Chang, D., Lindberg, E., & Winssinger, N.
- Affiliations
- University of Geneva (CH)
- Corresponding author
- Winssinger, Nicolas
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2017-01-24
- Volume
- 139
- Issue
- 4
- Pages
- 1444-1447
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W2579153981
- Cited by
- 72
- References
- 39
- Citations by year
- 2026: 5 · 2025: 4 · 2024: 7 · 2023: 4 · 2022: 9 · 2021: 10 · 2020: 12 · 2019: 6 · 2018: 6 · 2017: 9
- FWCI
- 3.69
- Type
- journal article
- Language
- en
- Open access
- Licence
- other-oa
- Locations
- Journal of the American Chemical Society (publishedVersion)
- PubMed (publishedVersion)
- Archive ouverte UNIGE (University of Geneva) (acceptedVersion, other-oa)
- Archive ouverte UNIGE (University of Geneva) (submittedVersion, cc-by)
- Funders
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (200020_157106)
- National Center of Competence in Research Chemical Biology
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Physical Sciences › Chemistry › Spectroscopy › Molecular Sensors and Ion Detection
- Keywords
- rate constants, photocatalytic reduction, ruthenium complexes, PNA probes, nucleic acid sensing, catalytic efficiency, signal amplification
- MeSH terms
- Nucleic Acids, Organometallic Compounds, Pyridinium Compounds, Ruthenium, Molecular Structure
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