RChIP-Seq: Chromatin-Associated RNA Sequencing in Developmentally Staged Mouse Testes

Methods Mol Biol. 2018:1832:169-184. doi: 10.1007/978-1-4939-8663-7_9.

Abstract

Chromatin is a dynamic macromolecular structure comprised of histones and a wealth of non-histone proteins. Recently, it has become clear that RNA is also an integral component of chromatin playing an important role in regulating its structure and function. Central to the understanding of RNA function is the ability to identify and genomically map interactions between chromatin components and RNA.Here, we describe a new method, RChIP-seq (RNA-associated-Chromatin-Immuno Precipitation followed by next-generation sequencing) that allows the identification of RNA species that are directly bound to specific components of chromatin in the mouse testis.

Keywords: CLIP; Chromatin; RNA-binding proteins; histone variant; next-generation sequencing.

MeSH terms

  • Animals
  • Chromatin Immunoprecipitation / methods*
  • Cross-Linking Reagents / chemistry
  • DNA, Complementary / genetics
  • Germ Cells / metabolism
  • High-Throughput Nucleotide Sequencing / methods*
  • Male
  • Mice, Inbred BALB C
  • Proteins / metabolism
  • RNA / metabolism
  • Sequence Analysis, RNA / methods*
  • Testis / growth & development*
  • Testis / metabolism*
  • Ultraviolet Rays

Substances

  • Cross-Linking Reagents
  • DNA, Complementary
  • Proteins
  • RNA