Enzymatic synthesis of reactive RNA probes containing squaramate-linked cytidine or adenosine for bioconjugations and cross-linking with lysine-containing peptides and proteins
2025

Creating RNA Probes for Protein Interaction Studies

publication Evidence: moderate

Author Information

Author(s): Ivana Ivancová, Tania Sánchez Quirante, Marek Ondruš, Radek Pohl, Marta Vlková, Eva Žilecká, Evžen Bouřa, Michal Hocek

Primary Institution: Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences

Hypothesis

Can squaramate-modified RNA probes be effectively synthesized for bioconjugation and cross-linking with lysine-containing proteins?

Conclusion

The study successfully synthesized squaramate-modified RNA probes that can specifically cross-link with lysine-containing peptides and proteins.

Supporting Evidence

  • The squaramate modification allows for specific cross-linking with lysine residues in proteins.
  • High conversion rates were achieved in bioconjugation reactions with excess amines.
  • RNA-protein cross-linking was confirmed through various analytical techniques.

Takeaway

Scientists made special RNA probes that can stick to proteins, helping them study how RNA and proteins interact.

Methodology

The study involved designing and synthesizing squaramate-modified ribonucleoside triphosphates and using them in in vitro transcription and primer extension reactions.

Limitations

The study did not achieve full conversion in all conjugation reactions, indicating potential inefficiencies.

Digital Object Identifier (DOI)

10.1038/s42004-024-01399-6

Want to read the original?

Access the complete publication on the publisher's website

View Original Publication