Multiplexed nanoflares: mRNA detection in live cells

Anal Chem. 2012 Feb 21;84(4):2062-6. doi: 10.1021/ac202648w. Epub 2012 Jan 30.

Abstract

We report the development of the multiplexed nanoflare, a nanoparticle agent that is capable of simultaneously detecting two distinct mRNA targets inside a living cell. These probes are spherical nucleic acid (SNA) gold nanoparticle (Au NP) conjugates consisting of densely packed and highly oriented oligonucleotide sequences, many of which are hybridized to a reporter with a distinct fluorophore label and each complementary to its corresponding mRNA target. When multiplexed nanoflares are exposed to their targets, they provide a sequence specific signal in both extra- and intracellular environments. Importantly, one of the targets can be used as an internal control, improving detection by accounting for cell-to-cell variations in nanoparticle uptake and background. Compared to single-component nanoflares, these structures allow one to determine more precisely relative mRNA levels in individual cells, improving cell sorting and quantification.

Publication types

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

MeSH terms

  • Flow Cytometry
  • Fluorescent Dyes*
  • Gold / chemistry*
  • Humans
  • Inhibitor of Apoptosis Proteins / genetics*
  • Metal Nanoparticles / chemistry*
  • Nanoparticles
  • Neoplasms / genetics
  • Nucleic Acid Hybridization
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Survivin
  • Tumor Cells, Cultured

Substances

  • BIRC5 protein, human
  • Fluorescent Dyes
  • Inhibitor of Apoptosis Proteins
  • RNA, Messenger
  • Survivin
  • Gold