Ultra-Specific G-Quadruplex-Colistin Interaction for Efficient Transcriptome-Wide G4 Mapping

Area of Science:

• Chemical Biology
• Genomics
• Molecular Biology

Background:

• G-quadruplexes (G4s) are complex nucleic acid structures with dynamic polymorphism, posing challenges for chemical biology interventions.
• Targeting G4s is crucial for understanding cellular processes and developing novel therapeutics.

Purpose of the Study:

• To investigate the interaction of small molecules with specific G4 structures.
• To develop a novel method for assessing the prevalence of RNA G-quadruplexes in the human transcriptome.

Main Methods:

• Exploited the specific interaction between the antibiotic colistin (COL) and parallel G-quadruplexes (G4s).
• Developed the COL-induced RNA G4 precipitation and sequencing (CoRP-seq) protocol, utilizing structure-specific aggregation and centrifugation for G4 isolation.
• Applied CoRP-seq to assess RNA G4 prevalence in human cell transcriptomes.

Main Results:

• Colistin (COL) selectively binds to parallel G-quadruplexes (G4s).
• This specific interaction induces G4/COL complex aggregation, allowing for separation via centrifugation.
• The CoRP-seq protocol demonstrated ultraspecificity, simplicity, and convenience for mapping RNA G4s.

Conclusions:

• The discovery of COL's specific G4 interaction provides a novel tool for G4 targeting.
• CoRP-seq offers a straightforward and highly specific method for RNA G-quadruplex profiling.
• This advancement addresses unmet needs in G4 mapping and the broader field of G4omics.