G-Quadruplex Visualization in Cells via Antibody and Fluorescence Probe

Area of Science:

• Molecular Biology
• Genetics
• Biochemistry

Background:

• G-quadruplexes (G4s) are noncanonical nucleic acid structures with crucial roles in gene regulation and disease across various organisms.
• Understanding G4 structure and function is vital for biological and pathological insights.
• Developing tools to visualize G4s in situ is essential for studying their biological relevance.

Purpose of the Study:

• To present three distinct protocols for visualizing G-quadruplexes (G4s) within cellular environments.
• To investigate G4 visualization in both uninfected and herpes simplex virus type 1 (HSV-1) infected cells.
• To assess the impact of a fluorescent G4 ligand on the G4-binding protein nucleolin.

Main Methods:

• Development and application of specific antibodies for G4 detection.
• Utilization of a fluorescent G4 ligand for G4 visualization.
• Microscopy techniques to observe G4s in cellular contexts, including HSV-1 infected cells.
• Analysis of the interaction between the fluorescent ligand, G4s, and nucleolin.

Main Results:

• Successful visualization of G-quadruplexes in cellular models using the developed protocols.
• Demonstration of G4 visualization in both normal and virus-infected cells (HSV-1).
• Observation of the fluorescent ligand's effect on nucleolin binding to G4 structures.

Conclusions:

• The presented protocols offer valuable tools for studying G-quadruplexes in biological systems.
• These methods facilitate the investigation of G4s' roles in cellular processes and viral infections.
• The study provides insights into the molecular interactions involving G4s, fluorescent ligands, and G4-binding proteins like nucleolin.