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Detection of TERRA R-Loops at Human Telomeres
Galina Glousker1, Rita Valador Fernandes1, Marianna Feretzaki1
1Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
Methods in Molecular Biology (Clifton, N.J.)
|June 15, 2022
Summary
Researchers developed a new method to detect R-loops, which are DNA-RNA structures, at telomeres. This technique helps understand how TERRA long noncoding RNA affects telomere maintenance and DNA repair.
Area of Science:
- Molecular Biology
- Genetics
- Epigenetics
Background:
- R-loops are nucleic acid structures comprising a DNA-RNA hybrid and a displaced DNA strand.
- The long noncoding RNA TERRA is known to form R-loops at telomeres.
- Telomeric R-loops influence chromatin composition and telomere maintenance, affecting DNA replication, repair, and telomerase activity.
Purpose of the Study:
- To describe a novel method for detecting R-loops specifically at telomeric regions.
- To investigate the role of TERRA-induced R-loops in telomere biology.
Main Methods:
- Immunoprecipitation using the S9.6 antibody, which recognizes R-loops.
- Detection of telomeric DNA via dot-blot hybridization with a radiolabeled telomeric probe.
- Quantitative PCR (qPCR) utilizing DNA primers specific for subtelomeric sequences.
Main Results:
- Successfully established a method to detect R-loops at telomeres.
- The described technique allows for the specific identification and analysis of telomeric R-loop structures.
Conclusions:
- The developed method provides a valuable tool for studying R-loops at telomeres.
- This technique will aid in understanding the functional implications of TERRA and R-loops in telomere maintenance and genomic stability.
Keywords:
DNA–RNA hybridsDRIPImmunoprecipitationR-loopsS9.6 monoclonal antibodyTERRA long noncoding RNATelomeresMore Related Videos
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