Counting R-loops: A Novel Quantification Method for Nuclear DNA:RNA Hybrids

R‐loops are three stranded nucleic acid structures that form during transcription; when the nascent strand of RNA invades the replication bubble, hybridizes with the DNA template strand and displaces the non‐template strand. For many years, these DNA:RNA hybrids were thought to only be byproducts of transcription. In recent years, however, it has been shown that R‐loops play many roles in diverse cellular processes, such as immunoglobulin class‐switching recombination, epigenetic regulation, and in DNA replication. R‐loop mis regulation can lead to DNA damage and genomic instability. Additionally, R‐loops have been shown to play a role in several neurodegenerative diseases. Although, our knowledge of R‐loops has grown in the last few years, detection and quantification of R‐loops can still be difficult and inaccurate. Here, we provide a novel method for quantification of nuclear R‐loops based on cellular fractionation and compare it with commonly used R‐loop quantification methods. Support or Funding Information Research was funded by HHMI Research Scholars program at NMSU, Cowboys for Cancer Research and the Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103451. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .