Detection of intra-brain cytoplasmic 1 (BC1) long noncoding RNA using graphene oxide-fluorescence beacon detector

Sci Rep. 2016 Mar 21:6:22552. doi: 10.1038/srep22552.

Abstract

Detection of cellular expression of long noncoding RNAs (lncRNAs) was elusive due to the ambiguity of exposure of their reactive sequences associated with their secondary/tertiary structures and dynamic binding of proteins around lncRNAs. Herein, we developed graphene-based detection techniques exploiting the quenching capability of graphene oxide (GO) flakes for fluorescent dye (FAM)-labeled single-stranded siRNAs and consequent un-quenching by their detachment from GO by matching lncRNAs. A brain cytoplasmic 1 (BC1) lncRNA expression was significantly decreased by a siRNA, siBC1-1. GO quenched the FAM-labeled siBC1-1 peptide nucleic acid (PNA) probe, and this quenching was recovered by BC1. While FAM-siBC1-1-PNA-GO complex transfected spontaneously mouse or human neural stem cells, fluorescence was recovered only in mouse cells having high BC1 expression. Fluorescent dye-labeled single-stranded RNA-GO probe could detect the reactive exposed nucleic acid sequence of a cytoplasmic lncRNA expressing in the cytoplasm, which strategy can be used as a detection method of lncRNA expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Biosensing Techniques
  • Cell Line
  • Fluorescent Dyes / chemistry*
  • Graphite / chemistry*
  • Humans
  • Inverted Repeat Sequences
  • Mice
  • Oxides / chemistry
  • RNA, Small Cytoplasmic / genetics*
  • RNA, Small Cytoplasmic / metabolism
  • Spectrometry, Fluorescence

Substances

  • BC1 RNA
  • Fluorescent Dyes
  • Oxides
  • RNA, Small Cytoplasmic
  • Graphite