Site-specific covalent modification of RNA guided by functionality-transfer oligodeoxynucleotides

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Efficient covalent modification of large RNA molecules is crucial for developing novel biological tools and therapeutic strategies.
• Existing methods often lack specificity or efficiency for complex RNA structures.

Purpose of the Study:

• To develop a general, site-specific method for RNA modification.
• To utilize a functional oligonucleotide (ODN) template for guiding RNA modification.
• To achieve cytosine-selective modification of RNA.

Main Methods:

• A functional ODN probe containing 6-thioguanosine was synthesized and modified with a methylenediketone derivative, creating an S-functionalized ODN.
• A functionality-transfer reaction was employed, transferring a modification from the sulfur atom of the ODN probe to the amino group of cytosine bases in the target RNA strand.
• The reaction was performed within DNA-RNA duplexes to leverage proximity effects for selectivity.

Main Results:

• Site-specific and cytosine-selective RNA modifications were successfully achieved using the developed method.
• The selectivity of the modification was attributed to the close proximity of the reactants within the DNA-RNA duplex structure.
• The method demonstrates potential for precise manipulation of large RNA molecules.

Conclusions:

• A novel, general method for site-specific RNA modification guided by a functional ODN template has been established.
• The developed technique allows for targeted covalent modification of cytosine bases in RNA with high selectivity.
• This approach holds promise for advancing RNA-based biological tools and therapeutic applications.