Three-Component Bioorthogonal Reactions on Cellular DNA and RNA

Area of Science:

• Chemical Biology
• Molecular Biology
• Nucleic Acid Chemistry

Background:

• Metabolic labeling of nucleic acids with bioorthogonal groups often suffers from low yields.
• Steric hindrance in copper-free click chemistry limits its efficiency with DNA and RNA.

Purpose of the Study:

• To develop a novel reagent for efficient bioorthogonal labeling of nucleic acids.
• To overcome limitations of existing click chemistry methods for DNA and RNA modification.

Main Methods:

• Synthesis of a cationic Sondheimer diyne derivative (DiMOC).
• Investigation of DiMOC's reactivity with azide groups in duplex DNA.
• Application of DiMOC for visualization of metabolically labeled nucleic acids in cells.

Main Results:

• DiMOC rapidly reacts with azide groups in duplex DNA, unlike other cyclooctynes.
• DiMOC facilitates strain-promoted cross-linking of two azide groups in DNA.
• Metabolic labeling and DiMOC treatment enabled visualization of newly synthesized DNA and RNA in whole cells.

Conclusions:

• DiMOC is a potent reagent for bioorthogonal modification of nucleic acids.
• This method provides a robust strategy for visualizing newly synthesized DNA and RNA.
• DiMOC expands the toolkit for chemical biology and molecular imaging.