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Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor

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Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor. / Kivrak, Ezgi; Pauzaite, Tekle; Copeland, Nikki et al.

In: Biosensors, Vol. 11, No. 1, 17, 08.01.2021.

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Kivrak E, Pauzaite T, Copeland N, Hardy J, Kara P, Firlak M et al. Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor. Biosensors. 2021 Jan 8;11(1):17. doi: 10.3390/bios11010017

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@article{11ca881854f642fc968ab9e1f566666f,
title = "Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor",
abstract = "The CRISPR-Cas9 system has facilitated the genetic modification of various model organisms and cell lines. The outcomes of any CRISPR-Cas9 assay should be investigated to ensure/improve the precision of genome engineering. In this study, carbon nanotube-modified disposable pencil graphite electrodes (CNT/PGEs) were used to develop a label-free electrochemical nanogenosensor for the detection of point mutations generated in the genome by using the CRISPR-Cas9 system. Carbodiimide chemistry was used to immobilize the 5′-aminohexyl-linked inosine-substituted probe on the surface of the sensor. After hybridization between the target sequence and probe at the sensor surface, guanine oxidation signals were monitored using differential pulse voltammetry (DPV). Optimization of the sensitivity of the nanogenoassay resulted in a lower detection limit of 213.7 nM. The nanogenosensor was highly specific for the detection of the precisely edited DNA sequence. This method allows for a rapid and easy investigation of the products of CRISPR-based gene editing and can be further developed to an array system for multiplex detection of different-gene editing outcomes.",
keywords = "CRISPR-Cas9, homology-directed repair (HDR), electrochemical genosensor, mutation detection, carbon nanotube-modified PGE",
author = "Ezgi Kivrak and Tekle Pauzaite and Nikki Copeland and John Hardy and Pinar Kara and Melike Firlak and Atike Yardimci and Selahattin Yilmaz and Fahreddin Palaz and Mehmet Ozsoz",
year = "2021",
month = jan,
day = "8",
doi = "10.3390/bios11010017",
language = "English",
volume = "11",
journal = "Biosensors",
issn = "2079-6374",
publisher = "MDPI - Open Access Publishing",
number = "1",

}

RIS

TY - JOUR

T1 - Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor

AU - Kivrak, Ezgi

AU - Pauzaite, Tekle

AU - Copeland, Nikki

AU - Hardy, John

AU - Kara, Pinar

AU - Firlak, Melike

AU - Yardimci, Atike

AU - Yilmaz, Selahattin

AU - Palaz, Fahreddin

AU - Ozsoz, Mehmet

PY - 2021/1/8

Y1 - 2021/1/8

N2 - The CRISPR-Cas9 system has facilitated the genetic modification of various model organisms and cell lines. The outcomes of any CRISPR-Cas9 assay should be investigated to ensure/improve the precision of genome engineering. In this study, carbon nanotube-modified disposable pencil graphite electrodes (CNT/PGEs) were used to develop a label-free electrochemical nanogenosensor for the detection of point mutations generated in the genome by using the CRISPR-Cas9 system. Carbodiimide chemistry was used to immobilize the 5′-aminohexyl-linked inosine-substituted probe on the surface of the sensor. After hybridization between the target sequence and probe at the sensor surface, guanine oxidation signals were monitored using differential pulse voltammetry (DPV). Optimization of the sensitivity of the nanogenoassay resulted in a lower detection limit of 213.7 nM. The nanogenosensor was highly specific for the detection of the precisely edited DNA sequence. This method allows for a rapid and easy investigation of the products of CRISPR-based gene editing and can be further developed to an array system for multiplex detection of different-gene editing outcomes.

AB - The CRISPR-Cas9 system has facilitated the genetic modification of various model organisms and cell lines. The outcomes of any CRISPR-Cas9 assay should be investigated to ensure/improve the precision of genome engineering. In this study, carbon nanotube-modified disposable pencil graphite electrodes (CNT/PGEs) were used to develop a label-free electrochemical nanogenosensor for the detection of point mutations generated in the genome by using the CRISPR-Cas9 system. Carbodiimide chemistry was used to immobilize the 5′-aminohexyl-linked inosine-substituted probe on the surface of the sensor. After hybridization between the target sequence and probe at the sensor surface, guanine oxidation signals were monitored using differential pulse voltammetry (DPV). Optimization of the sensitivity of the nanogenoassay resulted in a lower detection limit of 213.7 nM. The nanogenosensor was highly specific for the detection of the precisely edited DNA sequence. This method allows for a rapid and easy investigation of the products of CRISPR-based gene editing and can be further developed to an array system for multiplex detection of different-gene editing outcomes.

KW - CRISPR-Cas9

KW - homology-directed repair (HDR)

KW - electrochemical genosensor

KW - mutation detection

KW - carbon nanotube-modified PGE

U2 - 10.3390/bios11010017

DO - 10.3390/bios11010017

M3 - Journal article

VL - 11

JO - Biosensors

JF - Biosensors

SN - 2079-6374

IS - 1

M1 - 17

ER -