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Communication

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

1
Department of Analytical Chemistry, Faculty of Pharmacy, Ege University, Izmir 35100, Turkey
2
Department of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster LA1 4YQ, UK
3
Department of Chemistry, Faculty of Science and Technology, Lancaster University, Lancaster LA1 4YB, UK
4
Materials Science Institute, Lancaster University, Lancaster LA1 4YB, UK
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Department of Chemistry, Gebze Technical University, Gebze 41400, Turkey
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Department of Chemical Engineering, Izmir Institute of Technology, İzmir 35430, Turkey
7
Faculty of Medicine, Hacettepe University, Ankara 06100, Turkey
8
Faculty of Engineering, Near East University, Lefkoşa 99138, Turkey
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 28 November 2020 / Revised: 5 January 2021 / Accepted: 6 January 2021 / Published: 8 January 2021
(This article belongs to the Special Issue Application of CRISPR Cas Systems for Biosensing)
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. View Full-Text
Keywords: CRISPR-Cas9; homology-directed repair (HDR); electrochemical genosensor; mutation detection; carbon nanotube-modified PGE CRISPR-Cas9; homology-directed repair (HDR); electrochemical genosensor; mutation detection; carbon nanotube-modified PGE
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MDPI and ACS Style

Kivrak, E.; Pauzaite, T.; Copeland, N.A.; Hardy, J.G.; Kara, P.; Firlak, M.; Yardimci, A.I.; Yilmaz, S.; Palaz, F.; Ozsoz, M. Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor. Biosensors 2021, 11, 17. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11010017

AMA Style

Kivrak E, Pauzaite T, Copeland NA, Hardy JG, Kara P, Firlak M, Yardimci AI, Yilmaz S, Palaz F, Ozsoz M. Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor. Biosensors. 2021; 11(1):17. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11010017

Chicago/Turabian Style

Kivrak, Ezgi, Tekle Pauzaite, Nikki A. Copeland, John G. Hardy, Pinar Kara, Melike Firlak, Atike I. Yardimci, Selahattin Yilmaz, Fahreddin Palaz, and Mehmet Ozsoz. 2021. "Detection of CRISPR-Cas9-Mediated Mutations Using a Carbon Nanotube-Modified Electrochemical Genosensor" Biosensors 11, no. 1: 17. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11010017

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