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Updated: Feb 19, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Triplex forming ability of oligonucleotides containing 2'-O-methyl-2-thiouridine or 2-thiothymidine
Itaru Okamoto1, Kohji Seio, Mitsuo Sekine
1Department of Life Science, Tokyo Institute of Technology, Japan.
Modified oligonucleotides with 2-thiocarbonyl groups significantly stabilize parallel triplexes. These triplex forming oligonucleotides (TFOs) demonstrate high selectivity for matched DNA bases.
Area of Science:
- Biochemistry
- Oligonucleotide Chemistry
- Molecular Biology
Background:
- Triplex forming oligonucleotides (TFOs) are crucial for gene targeting and regulation.
- Modifications to TFOs can enhance their stability and binding properties.
- Understanding the structural basis of TFO stabilization is key to their therapeutic applications.
Purpose of the Study:
- To investigate the impact of 2'-O-methyl-2-thiouridine (s2Um) and 2-thiothymidine (s2T) modifications on TFO stability.
- To elucidate the role of the 2-thiocarbonyl group in stabilizing triplex structures.
- To assess the base recognition selectivity of modified TFOs.
Main Methods:
- Synthesis of oligonucleotides containing s2Um and s2T.
- UV melting temperature experiments to determine triplex stability.
- Analysis of base pairing selectivity.
Main Results:
- Oligonucleotides with s2Um or s2T modifications showed significantly enhanced stabilization of parallel triplexes.
- The 2-thiocarbonyl group's stacking effect was identified as the primary contributor to triplex stabilization.
- Modified TFOs exhibited high selectivity in distinguishing matched from mismatched Hoogsteen bases.
Conclusions:
- Incorporation of s2Um and s2T modifications effectively stabilizes parallel DNA triplexes.
- The 2-thiocarbonyl group is a key structural element for enhancing triplex stability.
- These modified TFOs offer promising selectivity for specific DNA targets, relevant for molecular biology applications.
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