Reversible Acylation of RNA Enables Activatable Biosensing

Area of Science:

• Chemical Biology
• RNA Therapeutics
• Molecular Biology

Background:

• Current RNA control methods often use UV light, posing phototoxicity risks in live cells.
• There is a need for safer, endogenous stimulus-responsive chemical tools for RNA manipulation.

Purpose of the Study:

• To develop a novel postsynthetic RNA acylation strategy for controllable RNA function.
• To demonstrate the application of this method in regulating CRISPR/Cas13a activity and DNAzyme catalysis.

Main Methods:

• Introduced boronate ester (BE) groups to 2'-hydroxyls of RNA via postsynthetic modification.
• Utilized hydrogen peroxide (H2O2) to trigger traceless release of 2'-hydroxyl via 1,6-elimination.
• Applied acylated crRNA for conditional CRISPR/Cas13a activity and acylated DNAzyme for reversible catalysis and cell imaging.

Main Results:

• Demonstrated conditional regulation of CRISPR/Cas13a activity for target RNA detection.
• Achieved reversible control of 8-17 DNAzyme activity through specific RNA acylation.
• Successfully applied the method for cell-selective imaging of metal ions in cancer cells.

Conclusions:

• Developed a simple, general, and cell-selective RNA acylation strategy responsive to endogenous stimuli.
• This method offers potential for constructing activatable RNA sensors and pre-RNA medicines.
• The approach provides a non-phototoxic alternative for controlling RNA properties and functions.