Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription-Reference-Cited by-全球学者库

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

Published:2023-03-31 Issue:4 Volume:13 Page:449
ISSN:2079-6374
Container-title:Biosensors
language:en
Short-container-title:Biosensors

Author:

Si Jingyi¹,Zhou Wei²,Fang Ying¹,Zhou Da¹,Gao Yifan¹,Yao Qunyan¹,Shen Xizhong¹,Zhu Changfeng¹^ORCID

Affiliation:

1. Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan University, Shanghai 200032, China

2. Department of School and Nutrition, Shanghai Yangpu District Center for Disease Control and Prevention, Shanghai 200090, China

Abstract

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.

Funder

Science and Technology Commission of Shanghai Municipality

Publisher

MDPI AG

Subject

Clinical Biochemistry,General Medicine,Analytical Chemistry,Biotechnology,Instrumentation,Biomedical Engineering,Engineering (miscellaneous)

Link

https://www.mdpi.com/2079-6374/13/4/449/pdf

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