R-PLA enables enhanced detection of R-loops in mammalian cells using proximity ligation - PubMed
4 hours ago
- #proximity ligation assay
- #DNA damage
- #R-loops
- R-PLA is a new method using proximity ligation with S9.6 and RNaseH1 antibodies to detect R-loops via fluorescence microscopy.
- It offers enhanced selectivity by requiring two antibodies in close proximity and increased sensitivity through signal amplification.
- Signals appear as foci, reducing background and easing quantification, allowing specific labeling at DNA damage sites, telomeres, and stalled replication forks.
- R-PLA detected increased nucleoplasmic R-loops after DNA damage induced by PARP inhibitors and ionizing radiation.
- The method is also compatible with flow cytometry for analyzing RNaseH1-associated R-loops.