Dataset Information

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.

SUBMITTER: Kivrak E

PROVIDER: S-EPMC7827051 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Publications

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

Kivrak Ezgi E Pauzaite Tekle T Copeland Nikki A NA Hardy John G JG Kara Pinar P Firlak Melike M Yardimci Atike I AI Yilmaz Selahattin S Palaz Fahreddin F Ozsoz Mehmet M

Biosensors 20210108 1

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 us ...[more]

PMID: 33429883

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Project description:Using a simple drop-casting technique, we successfully fabricated a sensitive electrochemical sensor based on the composite of reduced graphene oxide (RGO) and multiwalled carbon nanotubes (MWCNT) deposited on the surface of a glassy carbon electrode (GCE) for individual and simultaneous measurements of hydroquinone (HQ), dopamine (DA), and uric acid (UA). The nanocomposite of RGO/MWCNT was further characterized in terms of its structural properties, surface morphology, and topography using Raman, FT-IR spectroscopy, SEM, HRTEM, and AFM. Then, the proposed sensor for simultaneous measurement of HQ, DA, and UA based on RGO/MWCNT-modified GCE was investigated for its electrochemical behavior and electroanalytical performances using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). In addition, the composition ratio between RGO and MWCT was 1 : 1 showing the highest electrochemical response for simultaneous detection of HQ, DA, and UA. Owing to the synergistic effect between RGO and MWCNT leading to excellent conductivity properties, the proposed sensor exhibited improved electrochemical response at pH 7 toward the oxidation processes of HQ, DA, and UA on the surface of modified electrode. The proposed sensor demonstrated three well-defined anodic peaks of these analytes with their linear concentrations ranges of 3.0-150.0 μM for HQ, 4.0-100.0 μM for DA, and 2.0-70.0 μM for UA. The limit of detection values for the simultaneous detection of HQ, DA, and UA were found as follows 0.400 ± 0.014, 0.500 ± 0.006, and 0.300 ± 0.016 μM, respectively. The additional features of this proposed sensor are high reproducibility and stability for the simultaneous detection of HQ, DA, and UA with negligible interference effect from interferents such as Mg2+, K+, Cl-, ascorbic acid, and glucose. An acceptable percentage of recovery was also shown by this sensor for simultaneous measurements of HQ, DA, and UA using 6 samples of human urine. In summary, the RGO/MWCNT nanocomposite has been shown to be a promising platform for rapid, simple, and reliable determination of simultaneous measurements of HQ, DA, and UA in practical applications.

Dataset Information

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

Publications

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

Similar Datasets

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