Label-free highly sensitive detection of DNA approximate length and concentration by impedimetric CRISPR-dCas9 based biosensor technology
Makaleler > Tam Makale · 2021
Yazarlar: ZİHNİ ONUR UYGUN, Sevcan ATAY
BIOELECTROCHEMISTRY, SCI Expanded İndekslerine Giren Dergi, Uluslararası, 2021, cilt 140, ss 107812-107812
DOI: 10.1016/j.bioelechem.2021.107812
Özet
<p>In this study, we designed a CRISPR-dCas9-based biosensor with potential clinical use in glioblastoma</p><p>subtype discrimination through detection of isocitrate dehydrogenase R132H (IDH) mutation status.</p><p>The electrode was modified to detect mutant DNA cysteamine (Cys), PAMAM, dCas9 and sgRNA for</p><p>R132H mutations, respectively. The biosensor system we proposed was able not only to detect mutant</p><p>DNA, but also to measure the approximate length of DNA. Therefore, it can be considered that the biosensor</p><p>technology that we developed is novel in the field of DNA biosensors. Another superior capability of</p><p>the biosensor system is that it can simultaneously measure DNA concentration by electrochemical impedance</p><p>spectroscopy and DNA length by capacitive detection, which lowers the concentration-based falsepositive</p><p>signals. The calibration range was obtained between 100 fM and 1000 fM, LOD and LOQ were</p><p>also calculated as 33.96 fM and 102.91 fM respectively. Moreover, thanks to the sensitivity of the capacitive</p><p>detection, the biosensor was able to discriminate the same EIS signals of the 200 bp and 250 fM concentration</p><p>data and 1000 bp and 50 fM concentration data. In conclusion, the biosensor was capable of</p><p>detect target DNA and DNA length, simultaneously in minutes.</p>