dCas9-mediated nanoelectrokinetic direct detection of target gene for liquid biopsy

H Lee, J Choi, E Jeong, S Baek, HC Kim, JH Chae… - Nano …, 2018 - ACS Publications
Nano letters, 2018ACS Publications
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 …
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.
ACS Publications
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