Bioorthogonal masked acylating agents for proximity-dependent RNA labelling

Area of Science:

• Molecular Biology
• Cell Biology
• Biochemistry

Background:

• RNA localization is crucial for cellular functions and is tightly regulated.
• Existing proximity labeling methods are effective for proteins but limited for unbiased RNA mapping.

Purpose of the Study:

• To develop a novel method for unbiased spatial RNA mapping.
• To create a platform for analyzing RNA distribution in specific subcellular locations.

Main Methods:

• Development of novel α-alkoxy thioenol and chloroenol esters as potent acylating agents.
• Utilizing subcellular-localized bioorthogonal esterase for controlled probe activation.
• Implementing the bioorthogonal acylating agents for proximity labelling and sequencing (BAP-seq) for RNA analysis.

Main Results:

• Demonstrated the ability to map RNA distribution in membrane-bound and membrane-less organelles.
• Showcased the effectiveness of BAP-seq for precise spatial RNA analysis.
• Validated the controlled-release acylating agent chemistry for proximity labeling.

Conclusions:

• BAP-seq provides a robust and unbiased approach for mapping RNA localization.
• The developed method expands the capabilities of proximity labeling technologies for RNA studies.
• This platform offers a powerful tool to investigate cellular RNA organization.