Sequence-Specific Installation of Aryl Groups in RNA via DNA-Catalyst Conjugates

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Site-specific RNA modification is crucial for labeling, functionalization, and therapeutic development.
• Existing methods for RNA modification often face challenges in specificity and efficiency.

Purpose of the Study:

• To develop a novel catalytic strategy for sequence-specific RNA modification using DNA oligonucleotides.
• To demonstrate the utility of this method for RNA labeling and functionalization.

Main Methods:

• Utilized DNA oligonucleotides with a catalytic amine group for S_NAr arylation of RNA 2'-OH groups.
• Employed chloro-pyrimidine electrophiles and amino-DNA conjugates to form a transient ammonium aryl intermediate.
• Applied the strategy to label messenger RNA (mRNA) and achieve selective protein knockdown.

Main Results:

• Achieved high yields of heterocycle delivery to targeted RNA sites.
• Successfully fluorescently labeled mRNA at the polyA tail using an azide-containing aryl electrophile.
• Demonstrated selective in vitro arylation in mRNA coding regions, leading to targeted protein expression knockdown.

Conclusions:

• Developed a versatile, sequence-directed catalytic strategy for RNA modification and labeling.
• This approach offers a powerful tool for diverse applications in RNA research and therapeutics.
• The method enables precise functionalization of RNA molecules at specific locations.