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Nano Lett. 2018 Dec 12;18(12):7642-7650. doi: 10.1021/acs.nanolett.8b03224. Epub 2018 Nov 27.

dCas9-mediated Nanoelectrokinetic Direct Detection of Target Gene for Liquid Biopsy.

Author information

1
Department of Chemical and Biological Engineering , Jeju National University , Jeju , 63243 , Republic of Korea.
2
Center for Genome Engineering , Institute for Basic Science , Seoul 34047 , Republic of Korea.
3
Department of Internal Medicine , Seoul National University Hospital , Seoul 03080 , Republic of Korea.
4
Inter-university Semiconductor Research Center , Seoul National University , Seoul 08826 , Republic of Korea.

Abstract

The-state-of-the-art bio- and nanotechnology have opened up an avenue to noninvasive liquid biopsy for identifying diseases from biomolecules in bloodstream, especially DNA. In this work, we combined sequence-specific-labeling scheme using mutated clustered regularly interspaced short palindromic repeats associated protein 9 without endonuclease activity (CRISPR/dCas9) and ion concentration polarization (ICP) phenomenon as a mechanism to selectively preconcentrate targeted DNA molecules for rapid and direct detection. Theoretical analysis on ICP phenomenon figured out a critical mobility, elucidating two distinguishable concentrating behaviors near a nanojunction, a stacking and a propagating behavior. Through the modulation of the critical mobility to shift those behaviors, the C-C chemokine receptor type 5 ( CCR5) sequences were optically detected without PCR amplification. Conclusively, the proposed dCas9-mediated genetic detection methodology based on ICP would provide rapid and accurate micro/nanofluidic platform of liquid biopsies for disease diagnostics.

KEYWORDS:

Ion concentration polarization; dCas9; direct detection; liquid biopsy; micro/nanofluidics; selective preconcentration

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