MicroRNA-triggered dCas9 beacon for cancer detection

Date
2017
Journal Title
Journal ISSN
Volume Title
Publisher
University of Delaware
Abstract
Cancer is the second leading cause of death in the United States. Early detection has a major influence on receiving effective treatment and increasing the chance of survival. One such target for detection of cancer is microRNA. Deregulation of microRNA have been linked to a variety of cancers; which combined with their small size of 22 nucleotides makes them a popular target for cancer detection. ☐ In this work, an in vitro method for sensitive, rapid detection of cancer-relevant microRNA has been developed. The key of the assay relies on the ability of the type II catalytically inactive Cas9 protein, when complexed with its single-guide RNA, to recognize and bind to any designed double-stranded DNA target. The presence of a cancer-relevant microRNA triggers a series of toehold displacement reactions resulting in a fully assembled molecular beacon labeled by a fluorophore-quencher pair. Upon assembly of the beacon, a dCas9-sgRNA complex binds and displaces a quencher strand for fluorescence signal. This dCas9 beacon provides a microRNA detection limit at nanomolar concentrations.
Description
Keywords
Biological sciences, Applied sciences, Cancer, Cas9, MicroRNA, Molecular beacon, Sensor
Citation