Insight into lncRNA biology using hybridization capture analyses

Biochim Biophys Acta. 2016 Jan;1859(1):121-7. doi: 10.1016/j.bbagrm.2015.09.004. Epub 2015 Sep 14.

Abstract

Despite mounting evidence of the importance of large non-coding RNAs (lncRNAs) in biological regulation, we still know little about how these lncRNAs function. One approach to understand the function of lncRNAs is to biochemically purify endogenous lncRNAs from fixed cells using complementary oligonucleotides. These hybridization capture approaches can reveal the genomic localization of lncRNAs, as well as the proteins and RNAs with which they interact. To help researchers understand how these tools can uncover lncRNA function, this review discusses the considerations and influences of different parameters, (e.g., crosslinking reagents, oligonucleotide chemistry and hybridization conditions) and controls to avoid artifacts. By examining the application of these tools, this review will highlight the progress and pitfalls of studying lncRNAs using hybridization capture approaches.This article is part of a Special Issue entitled: Clues to long noncoding RNA taxonomy1, edited by Dr. Tetsuro Hirose and Dr. Shinichi Nakagawa.

Keywords: CHART; ChIRP; Chromatin; Formaldehyde; Genomic localization; RAP; lncRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Chromatin / genetics
  • Gene Expression Regulation
  • Genome*
  • Nucleic Acid Hybridization / genetics
  • Nucleic Acid Hybridization / methods*
  • Oligonucleotides, Antisense / chemistry
  • Oligonucleotides, Antisense / genetics*
  • RNA, Long Noncoding / chemistry
  • RNA, Long Noncoding / genetics*

Substances

  • Chromatin
  • Oligonucleotides, Antisense
  • RNA, Long Noncoding