Mutually Orthogonal Bioconjugation of Vinyl Nucleosides for RNA Metabolic Labeling

Area of Science:

• Bioconjugation Chemistry
• RNA Biology
• Chemical Biology

Background:

• Vinyl nucleosides offer versatile handles for bioconjugation.
• Orthogonal labeling strategies are crucial for complex biological studies.
• Current RNA labeling methods have limitations in studying nascent RNA populations.

Purpose of the Study:

• To develop an orthogonal conjugation strategy for vinyl nucleosides.
• To enable RNA modification for fluorescent cell imaging.
• To facilitate the study of nascent RNA expression.

Main Methods:

• Investigated selective reactivity of 5-vinyluridine with maleimide.
• Investigated selective reactivity of 2-vinyladenosine with tris(2-carboxyethyl)phosphine (TCEP).
• Utilized density functional theory (DFT) to study reaction mechanisms and orthogonality.
• Applied the developed chemistries for RNA modification and fluorescent cell imaging.

Main Results:

• Achieved orthogonal conjugation of 5-vinyluridine and 2-vinyladenosine.
• Demonstrated selective reactivity with maleimide and TCEP, respectively.
• Successfully modified RNA for fluorescent cell imaging.
• Enabled visualization of nascent RNA within different cellular populations.

Conclusions:

• The developed orthogonal conjugation strategy expands the utility of vinyl nucleosides.
• This method provides a powerful tool for RNA metabolic labeling and imaging.
• Facilitates advanced studies on nascent RNA dynamics and expression.