dCas9-mediated nanoelectrokinetic direct detection of target gene for liquid biopsy
- Lee, Hyomin; Choi, Jihye; Jeong, Euihwan; Baek, Seongho; Kim, Hee Chan; Chae, Jong-Hee; Koh, Youngil; Seo, Sang Woo; Kim, Jin-Soo; Kim, Sung Jae
- Issue Date
- Nano Letters, Vol.18 No.12, pp.7642-7650
- Ion concentration polarization; selective preconcentration; dCas9; liquid biopsy; micro/nanofluidics; direct detection
- 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.
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