LncRNA-Chromatin Pull-Down Using Biotin-Conjugated DNA Probes

Methods Mol Biol. 2022:2458:345-357. doi: 10.1007/978-1-0716-2140-0_19.

Abstract

Long noncoding RNAs (lncRNAs) are a class of RNA molecules that have been associated with several important biological processes and linked to numerous diseases. Due to their cell type- and tissue specific expression, lncRNAs are involved in a wide range of molecular pathways. To fully understand how a lncRNA is linked to a biological process, its mechanism of action needs to be uncovered. Nuclear retained lncRNAs have been described to modulate gene expression directly or indirectly by interacting with chromatin and associated factors. Described here is an RNA pull-down strategy, which enables the identification of chromatin regions directly bound by a lncRNA of interest. This method is an important step toward investigating how lncRNAs regulate gene expression and/or chromatin states.

Keywords: Chromatin; Pull-down assay; lncRNA.

MeSH terms

  • Biotin / genetics
  • Cell Nucleus / genetics
  • Cell Nucleus / metabolism
  • Chromatin* / genetics
  • DNA Probes
  • RNA, Long Noncoding* / genetics
  • RNA, Long Noncoding* / metabolism

Substances

  • Chromatin
  • DNA Probes
  • RNA, Long Noncoding
  • Biotin