In Vitro Silencing of lncRNAs Using LNA GapmeRs

Methods Mol Biol. 2021:2348:157-166. doi: 10.1007/978-1-0716-1581-2_10.

Abstract

Despite substantial advancements have been achieved in the identification of long noncoding RNA (lncRNA) molecules, many challenges still remain into their functional characterization. Loss-of-function approaches are needed to study oncogenic lncRNAs, which appear more difficult to knock down by RNA interference as compared to mRNAs. In this chapter, we present a protocol based on the use of a novel class of antisense oligonucleotides, named locked nucleic acid (LNA) GapmeRs, to inhibit the oncogenic lncRNA NEAT1 in multiple myeloma cells. Overall, this approach holds many advantages, including its possible independence from delivery reagents as well as the capability to knock down lncRNAs even in hard-to-transfect suspension cells, like hematopoietic cells.

Keywords: ASO; Electroporation; Gymnosis; LNA-GapmeR; Multiple myeloma cells; lncRNA; ncRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor
  • Cell Line, Tumor
  • Electroporation
  • Gene Knockdown Techniques*
  • Gene Silencing*
  • Humans
  • Oligonucleotides*
  • RNA Interference
  • RNA, Long Noncoding / genetics*
  • RNA, Messenger
  • Transfection

Substances

  • Biomarkers, Tumor
  • NEAT1 long non-coding RNA, human
  • Oligonucleotides
  • RNA, Long Noncoding
  • RNA, Messenger
  • locked nucleic acid