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Identification of RNA–DNA Hybrids Associated with R-Loops at the IgH Switch Sequence in Activated B Cells

  • Junghyun Lim
  • , Brice Laffleur
  • , Uttiya Basu*
  • , Kefei Yu
  • *Corresponding author for this work
  • Columbia University
  • University of Rennes
  • Michigan State University

Research output: Contribution to conferenceChapterpeer-review

Abstract

During transcription and replication, R-loops that contain RNA–DNA hybrids are generated across numerous genomic loci and contribute to many biological events. Using S9.6, a monoclonal antibody against RNA–DNA hybrids, accelerated the study of R-loop biology. An outpouring of recent studies has implicated various contributions of R-loop in physiological cellular functions. Earlier studies using nondenaturing sodium bisulfite probing also supported existence of DNA–RNA hybrids formation in mammalian cells. In activated B cells, RNA–DNA hybrids formation at IgH gene locus of B cells is crucial for class switch recombination that ensure the proper effector function of the antibody. Here, we describe the identification of R-loops associated with the IgH locus using RNA–DNA hybrid immunoprecipitation sequencing and nondenaturing sodium bisulfite probing. This will be helpful for future studies of R-loops status on whole genome as well as on IgH locus in B cells.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages55-66
Number of pages12
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2528
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • B cells
  • Class switch recombination
  • DRIP-seq
  • Nondenaturing sodium bisulfite probing

Quacquarelli Symonds(QS) Subject Topics

  • Biological Sciences

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