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

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...

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Detalles Bibliográficos
Autores principales: Si, Jingyi, Zhou, Wei, Fang, Ying, Zhou, Da, Gao, Yifan, Yao, Qunyan, Shen, Xizhong, Zhu, Changfeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10135927/
https://www.ncbi.nlm.nih.gov/pubmed/37185524
http://dx.doi.org/10.3390/bios13040449
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author Si, Jingyi
Zhou, Wei
Fang, Ying
Zhou, Da
Gao, Yifan
Yao, Qunyan
Shen, Xizhong
Zhu, Changfeng
author_facet Si, Jingyi
Zhou, Wei
Fang, Ying
Zhou, Da
Gao, Yifan
Yao, Qunyan
Shen, Xizhong
Zhu, Changfeng
author_sort Si, Jingyi
collection PubMed
description 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.
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spelling pubmed-101359272023-04-28 Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription Si, Jingyi Zhou, Wei Fang, Ying Zhou, Da Gao, Yifan Yao, Qunyan Shen, Xizhong Zhu, Changfeng Biosensors (Basel) Article 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. MDPI 2023-03-31 /pmc/articles/PMC10135927/ /pubmed/37185524 http://dx.doi.org/10.3390/bios13040449 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Si, Jingyi
Zhou, Wei
Fang, Ying
Zhou, Da
Gao, Yifan
Yao, Qunyan
Shen, Xizhong
Zhu, Changfeng
Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title_full Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title_fullStr Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title_full_unstemmed Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title_short Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription
title_sort label-free detection of t4 polynucleotide kinase activity and inhibition via malachite green aptamer generated from ligation-triggered transcription
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10135927/
https://www.ncbi.nlm.nih.gov/pubmed/37185524
http://dx.doi.org/10.3390/bios13040449
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