Conjugatable and Bioreduction Cleavable Linker for the 5'-Functionalization of Oligonucleotides

Area of Science:

• Chemical Biology
• Oligonucleotide Synthesis
• Bioconjugation

Background:

• Oligonucleotide modification is crucial for various applications.
• Efficient and cleavable linkers are needed for controlled oligonucleotide functionalization.

Purpose of the Study:

• To design and synthesize a novel linker for 5'-terminal oligonucleotide modification.
• To enable efficient conjugation and subsequent bioreductive cleavage of oligonucleotides.

Main Methods:

• Phosphoramidite reagent synthesis and incorporation into oligonucleotides via solid-phase synthesis.
• Copper-catalyzed azide-alkyne cycloaddition (CuAAC) for conjugating oligonucleotides to functional molecules.
• Nitroreductase/NADH system for bioreductive cleavage of the linker.

Main Results:

• A novel, efficient, conjugatable, and bioreduction-cleavable linker was successfully synthesized.
• The linker was incorporated at the 5'-terminus of oligonucleotides using standard solid-phase synthesis.
• Oligonucleotide conjugates were formed with diverse functional molecules via CuAAC reaction.
• The linker enabled the release of naked oligonucleotides through bioreduction.

Conclusions:

• The developed linker is a versatile tool for creating functionalized oligonucleotides.
• The linker facilitates controlled conjugation and efficient release of oligonucleotides.
• This methodology offers a robust platform for oligonucleotide-based applications.