Detection of DNA-RNA Hybrids In Vivo

Methods Mol Biol. 2018;1672:347-361. doi: 10.1007/978-1-4939-7306-4_24.

Abstract

DNA-RNA hybrids form naturally during essential cellular functions such as transcription and replication. However, they may be an important source of genome instability, a hallmark of cancer and genetic diseases. Detection of DNA-RNA hybrids in cells is becoming crucial to understand an increasing number of molecular biology processes in genome dynamics and function and to identify new factors and mechanisms responsible for disease in biomedical research. Here, we describe two different procedures for the reliable detection of DNA-RNA hybrids in the yeast Saccharomyces cerevisiae and in human cells: DNA-RNA Immunoprecipitation (DRIP) and Immunofluorescence.

Keywords: DNA-RNA hybrids; Genome instability; R loop; S9.6 antibody.

MeSH terms

  • Cell Line
  • DNA* / genetics
  • Fluorescent Antibody Technique
  • Genomic Instability
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Nucleic Acid Hybridization* / methods
  • RNA* / genetics
  • Saccharomyces cerevisiae / genetics

Substances

  • RNA
  • DNA