Please use this identifier to cite or link to this item:
https://hdl.handle.net/11147/11581
Title: | Detection of CRISPR-Cas9-Mediated mutations using a carbon nanotube-modified electrochemical genosensor | Authors: | Kıvrak, Ezgi Pauzaite, Tekle Copeland, Nikki A. Hardy, John G. Kara, Pınar Fırlak, Melike İnce Yardımcı, Atike Yılmaz, Selahattin Palaz, Fahreddin |
Keywords: | CRISPR-Cas9 Homology-directed repair (HDR) Electrochemical genosensor Mutation detection Carbon nanotube-modified PGE |
Publisher: | MDPI | 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. | URI: | https://doi.org/10.3390/bios11010017 https://hdl.handle.net/11147/11581 |
ISSN: | 2079-6374 |
Appears in Collections: | Chemical Engineering / Kimya Mühendisliği PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection |
Files in This Item:
File | Size | Format | |
---|---|---|---|
biosensors-11-00017.pdf | 2.16 MB | Adobe PDF | View/Open |
CORE Recommender
SCOPUSTM
Citations
10
checked on Nov 29, 2024
WEB OF SCIENCETM
Citations
9
checked on Oct 26, 2024
Page view(s)
258
checked on Dec 2, 2024
Download(s)
92
checked on Dec 2, 2024
Google ScholarTM
Check
Altmetric
Items in GCRIS Repository are protected by copyright, with all rights reserved, unless otherwise indicated.