Live-cell imaging unveils distinct R-loop populations with heterogeneous dynamics.
Robert M MartinMadalena R de AlmeidaEduardo GameiroSérgio Fernandes de AlmeidaPublished in: Nucleic acids research (2023)
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.
Keyphrases
- high resolution
- nucleic acid
- circulating tumor
- transcription factor
- induced apoptosis
- endothelial cells
- single molecule
- cell free
- binding protein
- protein protein
- amino acid
- genome wide
- quantum dots
- living cells
- signaling pathway
- fluorescence imaging
- dna methylation
- antibiotic resistance genes
- microbial community
- high density
- endoplasmic reticulum stress
- label free
- genetic diversity