State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China.
Anal Chim Acta. 2021 Dec 15;1188:339180. doi: 10.1016/j.aca.2021.339180. Epub 2021 Oct 18.
CRISPR-Cas12a system exhibits tremendous potential in accurate recognition and quantitation of nucleic acids and non-nucleic-acid targets thanks to the discovery of its cleavage capability toward single-stranded DNA (ssDNA). In this study, we developed an efficient electrochemiluminescence (ECL) sensing platform based on CRISPR-Cas12a for the analysis of adenosine triphosphate (ATP). In the presence of the target, the successful release of the DNA activator is specially recognized by Cas12a-crRNA duplex and activates the cleavage of ferrocene (Fc) labeled-ssDNA (Fc-ssDNA) modified on the cathode of bipolar electrode (BPE), resulting in a decrease of ECL intensity of [Ru(bpy)]/TPrA in the anodic cell of BPE. By means of the unique combination of Cas12a with ECL technique based on BPE, it can convert the recognition of target ATP into a detectable ECL signal. The detection limit of ATP was determined to be 0.48 nM under the optimal conditions. This work will expand the application of CRISPR-Cas detection system and propose a potential method for the analysis of non-nucleic-acid targets.
CRISPR-Cas12a 系统凭借其对单链 DNA（ssDNA）的切割能力，在准确识别和定量核酸和非核酸靶标方面显示出巨大的潜力。在本研究中，我们开发了一种基于 CRISPR-Cas12a 的高效电致化学发光（ECL）传感平台，用于分析三磷酸腺苷（ATP）。在靶标存在的情况下，DNA 激活剂的成功释放被 Cas12a-crRNA 双链体特异性识别，并激活双极电极（BPE）阴极上标记的二茂铁（Fc）标记的 ssDNA（Fc-ssDNA）的切割，导致 BPE 阳极池中的[Ru（bpy）] / TPrA 的 ECL 强度降低。通过 Cas12a 与基于 BPE 的 ECL 技术的独特结合，可以将靶标 ATP 的识别转化为可检测的 ECL 信号。在最佳条件下，ATP 的检测限确定为 0.48 nM。这项工作将扩展 CRISPR-Cas 检测系统的应用，并为非核酸靶标的分析提出一种潜在的方法。
