| Yazarlar |
Doç. Dr. Zihni Onur UYGUN
Kafkas Üniversitesi, Türkiye |
Levent Yeniay
Ege Üniversitesi, Türkiye |
|
Ferhan Girgin Sağın
Ege Üniversitesi, Türkiye |
| Özet |
| Label-free biosensors which can be integrated into lab-on-a-chip platforms have the advantage of using small volumes for rapid and inexpensive measurements contrary to label-based technologies which are often more costly and time-consuming. In this study, graphene oxide screen printed electrodes (GPHOXE) were modified by deactivated Cas9 (dCas9) proteins and synthetic guide RNA (sgRNA) as the biorecognition receptor for label-free detection of circulating tumor DNAs (ctDNA). This was achieved by detection of a tumor related mutation (PIK3CA exon 9 mutation) via sequence-specific recognition followed by electrochemical impedance spectroscopy (EIS) analysis. The biosensor showed high specificity as there was no impedance signal for other ctDNA sequences, even the single nucleotide mismatch. dCas9-sgRNA modified biosensor demonstrated linear detection limits between 2 and 20 nM for 120 bp ctDNA's in 40 s. The calibration curve showed good linearity, LOD was calculated as 0.65 nM and LOQ was calculated as 1.92 nM. Selectivity and repeatability studies were carried out in real blood samples and the recovery was higher than 96%. In conclusion, dCas9-sgRNA was effectively immobilized and optimized on GPHOXE as the selective biorecognition receptor of this ultrafast impedimetric biosensor. The CRISPR-dCas9 powered impedimetric system showed good selectivity, high repeatability and good recovery properties. This is the first literature to report the use of CRISPR/Cas technology as a label-free tool that can be used in an impedimetric system for detection of ctDNA's. |
| Anahtar Kelimeler |
| Impedimetric biosensor,Circulating tumor DNA,Chronoimpedance,CRISPR/Cas9 technology,Deactivated Cas9,Label-free analysis,Electrochemical impedance spectroscopy,Liquid biopsy,Cancer diagnosis |
| Makale Türü | Özgün Makale |
| Makale Alt Türü | SSCI, AHCI, SCI, SCI-Exp dergilerinde yayımlanan tam makale |
| Dergi Adı | Analytica Chimica Acta |
| Dergi ISSN | 0003-2670 |
| Dergi Tarandığı Indeksler | SCI-Expanded |
| Dergi Grubu | Q1 |
| Makale Dili | İngilizce |
| Basım Tarihi | 07-2020 |
| Cilt No | 1121 |
| Sayfalar | 35 / 41 |
| Doi Numarası | 10.1016/j.aca.2020.04.009 |
| Makale Linki | http://dx.doi.org/10.1016/j.aca.2020.04.009 |
| Atıf Sayıları | |
| WoS | 75 |
| SCOPUS | 80 |
| Google Scholar | 92 |
