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@article{d0d994065b8f429289c0cf02de707903,
title = "Detection of CRISPR-dCas9 on DNA with Solid-State Nanopores",
abstract = "Solid-state nanopores have emerged as promising platforms for biosensing including diagnostics for disease detection. Here we show nanopore experiments that detect CRISPR-dCas9, a sequence-specific RNA-guided protein system that specifically binds to a target DNA sequence. While CRISPR-Cas9 is acclaimed for its gene editing potential, the CRISPR-dCas9 variant employed here does not cut DNA but instead remains tightly bound at a user-defined binding site, thus providing an excellent target for biosensing. In our nanopore experiments, we observe the CRISPR-dCas9 proteins as local spikes that appear on top of the ionic current blockade signal of DNA molecules that translocate through the nanopore. The proteins exhibit a pronounced blockade signal that allows for facile identification of the targeted sequence. Even at the high salt conditions (1 M LiCl) required for nanopore experiments, dCas9 proteins are found to remain stably bound. The binding position of the target sequence can be read from the spike position along the DNA signal. We anticipate applications of this nanopore-based CRISPR-dCas9 biosensing approach in DNA-typing based diagnostics such as quick disease-strain identification, antibiotic-resistance detection, and genome typing.",
keywords = "biosensing, CRISPR-Cas9, diagnostics, Nanopores",
author = "Wayne Yang and Laura Restrepo-P{\'e}rez and Michel Bengtson and Heerema, {Stephanie J.} and Anthony Birnie and {Van Der Torre}, Jaco and Cees Dekker",
year = "2018",
doi = "10.1021/acs.nanolett.8b02968",
language = "English",
journal = "Nano Letters: a journal dedicated to nanoscience and nanotechnology",
issn = "1530-6984",
publisher = "American Chemical Society",

}

RIS

TY - JOUR

T1 - Detection of CRISPR-dCas9 on DNA with Solid-State Nanopores

AU - Yang, Wayne

AU - Restrepo-Pérez, Laura

AU - Bengtson, Michel

AU - Heerema, Stephanie J.

AU - Birnie, Anthony

AU - Van Der Torre, Jaco

AU - Dekker, Cees

PY - 2018

Y1 - 2018

N2 - Solid-state nanopores have emerged as promising platforms for biosensing including diagnostics for disease detection. Here we show nanopore experiments that detect CRISPR-dCas9, a sequence-specific RNA-guided protein system that specifically binds to a target DNA sequence. While CRISPR-Cas9 is acclaimed for its gene editing potential, the CRISPR-dCas9 variant employed here does not cut DNA but instead remains tightly bound at a user-defined binding site, thus providing an excellent target for biosensing. In our nanopore experiments, we observe the CRISPR-dCas9 proteins as local spikes that appear on top of the ionic current blockade signal of DNA molecules that translocate through the nanopore. The proteins exhibit a pronounced blockade signal that allows for facile identification of the targeted sequence. Even at the high salt conditions (1 M LiCl) required for nanopore experiments, dCas9 proteins are found to remain stably bound. The binding position of the target sequence can be read from the spike position along the DNA signal. We anticipate applications of this nanopore-based CRISPR-dCas9 biosensing approach in DNA-typing based diagnostics such as quick disease-strain identification, antibiotic-resistance detection, and genome typing.

AB - Solid-state nanopores have emerged as promising platforms for biosensing including diagnostics for disease detection. Here we show nanopore experiments that detect CRISPR-dCas9, a sequence-specific RNA-guided protein system that specifically binds to a target DNA sequence. While CRISPR-Cas9 is acclaimed for its gene editing potential, the CRISPR-dCas9 variant employed here does not cut DNA but instead remains tightly bound at a user-defined binding site, thus providing an excellent target for biosensing. In our nanopore experiments, we observe the CRISPR-dCas9 proteins as local spikes that appear on top of the ionic current blockade signal of DNA molecules that translocate through the nanopore. The proteins exhibit a pronounced blockade signal that allows for facile identification of the targeted sequence. Even at the high salt conditions (1 M LiCl) required for nanopore experiments, dCas9 proteins are found to remain stably bound. The binding position of the target sequence can be read from the spike position along the DNA signal. We anticipate applications of this nanopore-based CRISPR-dCas9 biosensing approach in DNA-typing based diagnostics such as quick disease-strain identification, antibiotic-resistance detection, and genome typing.

KW - biosensing

KW - CRISPR-Cas9

KW - diagnostics

KW - Nanopores

UR - http://resolver.tudelft.nl/uuid:d0d99406-5b8f-4292-89c0-cf02de707903

UR - http://www.scopus.com/inward/record.url?scp=85053617399&partnerID=8YFLogxK

U2 - 10.1021/acs.nanolett.8b02968

DO - 10.1021/acs.nanolett.8b02968

M3 - Article

JO - Nano Letters: a journal dedicated to nanoscience and nanotechnology

T2 - Nano Letters: a journal dedicated to nanoscience and nanotechnology

JF - Nano Letters: a journal dedicated to nanoscience and nanotechnology

SN - 1530-6984

ER -

ID: 46905696