Imperial College London
Alternate

Professor Molly Stevens

Faculty of Engineering, Department of Materials

Professor of Biomedical Materials and Regenerative Medicine
 
 
 
//

Contact

 

+44 (0)20 7594 6804m.stevens

 
 
//

Location

 

208Royal School of MinesSouth Kensington Campus

//

Summary

 

Publications

Citation

BibTex format

@article{Kim:2016:10.1021/acssensors.6b00601,
author = {Kim, E and Howes, PD and Crowder, SW and Stevens, MM},
doi = {10.1021/acssensors.6b00601},
journal = {ACS Sensors},
pages = {111--118},
title = {Multi-Amplified Sensing of MicroRNA by a Small DNA Fragment-Driven Enzymatic Cascade Reaction},
url = {http://dx.doi.org/10.1021/acssensors.6b00601},
volume = {2},
year = {2016}
}
Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Combining technological developments such asnanomaterials, DNA nanotechnology, and functional enzymeshas great potential to facilitate next generation high performancemolecular diagnostic systems. In this work, we describe amicroRNA (miRNA) detection assay that combines targetrecycling and isothermal amplification in an elegantly designedenzyme-mediated cascade reaction. Target recycling is drivenby the action of duplex-specific nuclease (DSN), resulting inhighly amplified translation of input miRNA to short outputDNA fragments. These fragments act as highly specificinitiators of rolling circle amplification (RCA), an isothermalreaction that outputs a large volume of polymeric DNAzymesper initiator, and finally a fluorogenic output signal. Based oncareful electrophoretic analysis we observed that the DSN produces ca. 10 nt DNA fragments from DNA/miRNA duplexes,regardless of the length of DNA strands. Target recycling yielded ca. 5 orders of magnitude amplification through the DSNassistedrecycling system on magnetic particles, and the RCA yielded a further 2 orders of magnitude. The final assay exhibited alimit of detection of 1.8 fM of miRNA spiked into 20% human serum, and showed excellent selectivity for miR-21 versus singlebase-mismatched sequences and other cancer-related miRNAs. The developed assay was further employed to determine accurateamounts of miR-21 in total RNA samples extracted from human cancer cell lines and normal cells, confirming the applicability ofthe assay for direct and absolute quantification of mature specific miRNA in real biological samples.
AU  - Kim,E
AU  - Howes,PD
AU  - Crowder,SW
AU  - Stevens,MM
DO  - 10.1021/acssensors.6b00601
EP  - 118
PY  - 2016///
SN  - 2379-3694
SP  - 111
TI  - Multi-Amplified Sensing of MicroRNA by a Small DNA Fragment-Driven Enzymatic Cascade Reaction
T2  - ACS Sensors
UR  - http://dx.doi.org/10.1021/acssensors.6b00601
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000393088300015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - http://hdl.handle.net/10044/1/48481
VL  - 2
ER  -
Download