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Updated: Jul 20, 2026
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A G-quadruplex DNA-affinity Approach for Purification of Enzymatically Active G4 Resolvase1
Published on: March 18, 2017
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Protocol for cellular RNA G-quadruplex profiling using G4RP.v2
Jérémie Mitteaux1, David Monchaud1
1Institut de Chimie Moléculaire de l'Université de Bourgogne (ICMUB), CNRS UMR 6302, 9 Avenue Alain Savary, 21078 Dijon, France.
STAR Protocols
|December 11, 2024
Summary
Researchers improved a protocol to isolate G-quadruplexes (G4s) in RNA from human cancer cells. This method aids in understanding the biological roles of these important nucleic acid structures.
Area of Science:
- Molecular Biology
- Genomics
- Biochemistry
Background:
- G-quadruplexes (G4s) are higher-order nucleic acid structures crucial in various biological processes.
- Identifying and studying G4s in RNA within human cells is essential for understanding their functions.
- Existing methods for G4 isolation present challenges in efficiency and specificity.
Purpose of the Study:
- To present an optimized G4-RNA precipitation (G4RP) protocol for robust identification of RNA G4s in human cancer cells.
- To refine the methodology for isolating and analyzing G4 structures in RNA.
Main Methods:
- The improved protocol involves specific cell treatment and lysis procedures.
- Chemoprecipitation of G4s is performed using TASQ tools.
- Quantitative reverse-transcription PCR (RT-qPCR) is employed for quantification and analysis.
Main Results:
- The enhanced G4RP protocol provides a reliable method for isolating RNA G4s.
- The protocol includes critical go/no-go steps for quality control.
- Detailed steps for analysis ensure accurate data interpretation.
Conclusions:
- The improved G4RP protocol is a valuable tool for researchers studying RNA G4 biology in human cancer.
- This refined technique facilitates the discovery of the biological roles of G4 structures in RNA.
- The protocol aids in advancing the understanding of nucleic acid structures in cellular processes.
