Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription

作者：Jingyi Si, Wei Zhou, Ying Fang, Da Zhou, Yifan Gao, Qunyan Yao, Xizhong Shen, Changfeng Zhu · 发表于：Biosensors · 年份：2023 · DOI：10.3390/bios13040449 · 被引用次数：8 · 研究领域：Advanced biosensing and bioanalysis techniques、RNA Interference and Gene Delivery、DNA and Nucleic Acid Chemistry

Polynucleotide kinase (PNK) is a key enzyme that is necessary for ligation-based DNA repair. The activity assay and inhibitor screening for PNK may contribute to the prediction and improvement of tumor treatment sensitivity, respectively. Herein, we developed a simple, low-background, and label-free method for both T4 PNK activity detection and inhibitor screening by combining a designed ligation-triggered T7 transcriptional amplification system and a crafty light-up malachite green aptamer. Moreover, this method successfully detected PNK activity in the complex biological matrix with satisfactory outcomes, indicating its great potential in clinical practice.