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

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Aptamer Malachite green Chemistry Ligation DNA Proximity ligation assay Polynucleotide Biochemistry Molecular biology Cell biology Biology Receptor Adsorption Organic chemistry

Jingyi Si , Wei Zhou , Ying Fang , Da Zhou , Yifan Gao , Qunyan Yao , Xizhong Shen , Changfeng Zhu ·

YOU? · · 2023 · Open Access · · DOI: https://doi.org/10.3390/bios13040449 · OA: W4362587153

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.