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Detection of miRNA expression in intact cells using activatable sensor oligonucleotides.

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.

SUBMITTER: Yoo B

PROVIDER: S-EPMC3943878 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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Detection of miRNA expression in intact cells using activatable sensor oligonucleotides.

Yoo Byunghee B Kavishwar Amol A Ghosh Subrata K SK Barteneva Natalie N Yigit Mehmet V MV Moore Anna A Medarova Zdravka Z

Chemistry & biology 20140116 2

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 s ...[more]

PMID: 24440078

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