Rapid electronic detection of probe-specific microRNAs using thin nanopore sensors.
Small RNA molecules have an important role in gene regulation and RNA silencing therapy, but it is challenging to detect these molecules without the use of time-consuming radioactive labelling assays or error-prone amplification methods. Here, we present a platform for the rapid electronic detection of probe-hybridized microRNAs from cellular RNA. In this platform, a target microRNA is first hybridized to a probe. This probe:microRNA duplex is then enriched through binding to the viral protein p19. Finally, the abundance of the duplex is quantified using a nanopore. Reducing the thickness of the membrane containing the nanopore to 6 nm leads to increased signal amplitudes from biomolecules, and reducing the diameter of the nanopore to 3 nm allows the detection and discrimination of small nucleic acids based on differences in their physical dimensions. We demonstrate the potential of this approach by detecting picogram levels of a liver-specific miRNA from rat liver RNA.
Wanunu
Meni
M
Department of Physics and Astronomy, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Dadosh
Tali
T
Ray
Vishva
V
Jin
Jingmin
J
McReynolds
Larry
L
Drndić
Marija
M
D000818:Animals; D004247:DNA / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D008099:Liver / Q000737:chemistry; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D058608:Nanopores; D036103:Nanotechnology / Q000379:methods*; D015341:Nucleic Acid Probes / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D011119:Polynucleotides / Q000737:chemistry; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D051381:Rats; D014764:Viral Proteins
D035683:MicroRNAs; D015341:Nucleic Acid Probes; D011119:Polynucleotides; D014764:Viral Proteins; D012313:RNA; D004247:DNA