Sprecher
Beschreibung
The applicability of CRISPR-Cas has gone beyond genome editing and has found expansive use in molecular diagnostics. While there have been many fluorescent sensors, electrochemical sensors for CRISPR-Cas are now rapidly gaining attention as they reduce the need for expensive instruments, are low-cost, and can be used in low-resource settings, satisfying the requirements for an ideal point-of-care testing (POCT) device [1].
Conventional electrochemical sensors focus on a signal-off strategy, which limits detection range and sensitivity. In this study, we present the initial development of a novel electrochemical sensing platform based on nanoelectrode arrays (NEAs) [2] coupled with a signal-on strategy.
NEAs are fabricated by forming arrays of nanoholes in a thin polycarbonate (PC) film deposited on a conductive substrate through Thermal Nanoimprinting Lithography (TNIL). The presence of carboxylic acid groups on PC enables covalent functionalization with oligonucleotide probes, forming the basis of a biosensing interface [3]. Surface activation methods were evaluated, with NaOH treatment demonstrating superior performance compared to UV/Ozone (UVO). Conditions for covalent DNA immobilization were optimized, with improved functionalization observed at basic pH 8 and optimal DNA concentration of 0.5 µM [4]. Initial CRISPR-Cas assay results on PC substrates are also presented. Future work will explore the use of NEAs in electrochemical sensing and the development of microfluidic devices for single and multiplexed CRISPR-Cas assays. The combination of NEAs and CRISPR-Cas holds strong potential to advance molecular diagnostics for clinical and environmental applications.
References
[1] Y. Dai, R. A Somoza, L. Wang, J. F. Welter, Y. Li, A. I Caplan, C. C. Liu, Angew. Chem. Int. Ed. 2019, 58, 17399.
[2] D.W.M Arrigan, Analyst. 2004, 129, 1157-1165
[3] A. Zanut, A. Cian, N. Cefarin, A. Pozzato, M. Tormen, Biosensors. 2020, 10(8), 90
[4] S. Fortunati, I. Vasini, M.Giannetto, M. Mattarozzi, A.Porchetta, A.Bertucci, M. Careri, Anal. Chem. 2022, 94, 5075-5083
