Detection of miRNA expression in intact cells using activatable sensor oligonucleotides

Byunghee Yoo, Amol Kavishwar, Subrata K. Ghosh, Natalie Barteneva, Mehmet V. Yigit, Anna Moore, Zdravka Medarova

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We describe a technology for the profiling of miRNA expression in intact cells. The technology is based on sensor oligonucleotides that are cleavable, completely complementary to a target miRNA, and dual-labeled with a fluorescent dye and a quencher. Upon entering the cell, the sensor oligonucleotide binds its specific miRNA target through complementary base-pairing. This triggers assembly of the endogenous RNA Induced Silencing Complex (RISC) around the miRNA-sensor duplex and cleavage of the sensor oligonucleotide, resulting in separation between the dye and quencher, and a fluorescence turn-on. In the presented feasibility studies, we focus on a specific miRNA (miR-10b) implicated in breast cancer metastasis. Using a human breast adenocarcinoma cell line, we illustrate the application of this technology for miRNA detection with nanomolar sensitivity in both a cell-free system and intact cells.

Original languageEnglish
Pages (from-to)199-204
Number of pages6
JournalChemistry and Biology
Volume21
Issue number2
DOIs
Publication statusPublished - Feb 20 2014
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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