Designing Reversible Photoswitching Azobenzene-Modified Nucleotide for Controlling Biological Function

Area of Science:

• Chemical Biology
• Molecular Biology
• RNA Therapeutics

Background:

• RNA molecules are crucial for cellular functions and disease processes.
• Controlling RNA structure and function with light offers a powerful method for biological regulation and therapeutic development.

Purpose of the Study:

• To design and synthesize azobenzene-modified cytidine phosphoramidite for creating photoswitchable oligoribonucleotides.
• To investigate the reversible photoisomerization and impact of this modification on RNA structure and function.
• To demonstrate light-controlled regulation of the reverse transcription process using modified RNA.

Main Methods:

• Synthesis of azobenzene-modified cytidine phosphoramidite and oligoribonucleotides.
• Validation of photoisomerization using UV melting experiments and all-atom molecular dynamics (MD) simulations.
• Demonstration of light-induced switching of the reverse transcription (RT) process with various RT enzymes.

Main Results:

• Successful synthesis and validation of reversible photoisomerization in modified ribonucleosides and oligoribonucleotides.
• MD simulations and UV melting experiments revealed the impact of the cytidine modification.
• Demonstrated optical control over the RT process, including HIV reverse transcriptase-mediated RT, elucidated by MD simulation.

Conclusions:

• A novel chemical biology toolset for reversible optical control of RNA structures and functions has been developed.
• This technology holds promise for advancing gene regulation strategies and the development of novel RNA-based therapeutics.