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Updated: Jul 13, 2025

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Live-cell imaging unveils distinct R-loop populations with heterogeneous dynamics.

Robert M Martin1, Madalena R de Almeida1, Eduardo Gameiro1

  • 1Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, Lisboa, Portugal.

Nucleic Acids Research
|October 11, 2023
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Summary
This summary is machine-generated.

We created RHINO, a new sensor for visualizing RNA:DNA hybrids (R-loops) in live cells. This tool reveals how R-loop dynamics differ across cellular regions, offering insights into their roles in health and disease.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • R-loops, which are RNA:DNA hybrids, form during transcription and are implicated in genome stability and disease.
  • Existing methods for studying R-loops often lack the specificity or real-time imaging capabilities needed for dynamic cellular studies.

Purpose of the Study:

  • To develop a novel, genetically encoded sensor for live-cell imaging of R-loops.
  • To characterize the abundance, dynamics, and regional variations of R-loops in live cells.

Main Methods:

  • Development of RHINO, a sensor comprising RNA:DNA hybrid binding domains of human RNase H1 fused to a fluorescent protein.
  • Utilizing RHINO for high-specificity and sensitivity measurement of R-loop dynamics in live cells.
  • Kinetic analysis of R-loops at nucleoli, telomeres, and protein-coding genes.

Main Results:

  • RHINO enables precise live-cell imaging of R-loops with high specificity and sensitivity.
  • Significant variations in R-loop dynamics were observed across nucleoli, telomeres, and protein-coding genes.
  • A kinetic framework for R-loop behavior in different genomic contexts was established.

Conclusions:

  • RHINO is a powerful tool for studying R-loop biology in real-time within live cells.
  • R-loop dynamics are context-dependent, suggesting specialized roles in various chromosomal regions.
  • Understanding R-loop dynamics is crucial for elucidating their contribution to cellular processes and disease pathogenesis.