Custom long non-coding RNA capture enhances detection sensitivity in different human sample types

RNA Biol. 2021 Oct 15;18(sup1):215-222. doi: 10.1080/15476286.2021.1971438. Epub 2021 Sep 1.

Abstract

Long non-coding RNAs (lncRNAs) are a heterogeneous group of transcripts that lack protein coding potential and display regulatory functions in various cellular processes. As a result of their cell- and cancer-specific expression patterns, lncRNAs have emerged as potential diagnostic and therapeutic targets. The accurate characterization of lncRNAs in bulk transcriptome data remains challenging due to their low abundance compared to protein coding genes. To tackle this issue, we describe a unique short-read custom lncRNA capture sequencing approach that relies on a comprehensive set of 565,878 capture probes for 49,372 human lncRNA genes. This custom lncRNA capture approach was evaluated on various sample types ranging from artificial high-quality RNA mixtures to more challenging formalin-fixed paraffin-embedded tissue and biofluid material. The custom enrichment approach allows the detection of a more diverse repertoire of lncRNAs, with better reproducibility and higher coverage compared to classic total RNA-sequencing.

Keywords: FFPE; RNA abundance; RNA expression; RNA sequencing; biofluid; lncRNA; lncRNome; probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Body Fluids / chemistry*
  • Body Fluids / metabolism
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Paraffin Embedding / methods*
  • RNA, Long Noncoding / analysis*
  • RNA, Long Noncoding / isolation & purification*
  • RNA, Long Noncoding / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • RNA, Long Noncoding

Grants and funding

This work was supported by 'Fonds Wetenschappelijk Onderzoek' [1226821N and 1S07416N to C.E.; 1133120N to E.H.; 11C1621N to A.M.] and the Special Research Fund (BOF) of Ghent University [BOF.DOC.2019.0047.01]. This research is partly funded by ‘RNA-MAGIC’ and ‘LNCCA’ Concerted Research Actions of Ghent University (BOF19/GOA/008 and BOF16/GOA/023), by ‘Kom Op Tegen Kanker’ (Stand Up To Cancer, the Flemish cancer society) and by the Foundation Against Cancer.