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

• Autor: Martin, Robert M ; de Almeida, Madalena R ; Gameiro, Eduardo ; de Almeida, Sérgio F
• Assuntos: Gene regulation, Chromatin and Epigenetics
• É parte de: Nucleic acids research, 2023-11, Vol.51 (20), p.11010-11023
• Notas: ObjectType-Article-1
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• Descrição: Abstract We have developed RHINO, a genetically encoded sensor that selectively binds RNA:DNA hybrids enabling live-cell imaging of cellular R-loops. RHINO comprises a tandem array of three copies of the RNA:DNA hybrid binding domain of human RNase H1 connected by optimized linker segments and fused to a fluorescent protein. This tool allows the measurement of R-loop abundance and dynamics in live cells with high specificity and sensitivity. Using RHINO, we provide a kinetic framework for R-loops at nucleoli, telomeres and protein-coding genes. Our findings demonstrate that R-loop dynamics vary significantly across these regions, potentially reflecting the distinct roles R-loops play in different chromosomal contexts. RHINO is a powerful tool for investigating the role of R-loops in cellular processes and their contribution to disease development and progression. Graphical Abstract Graphical Abstract
  
• Editor: Oxford University Press
  
• Idioma: Inglês