A fluorescence strategy for circRNA quantification in tumor cells based on T7 nuclease-assisted cycling enzymatic amplification

被引：0

作者：

Qu, Hongke ^{[1
,2
]}

Chen, Mingjian ^{[2
]}

Ge, Junshang ^{[2
]}

Zhang, Xiangyan ^{[6
]}

He, Shuyi ^{[2
]}

Xiong, Fang ^{[3
]}

Yan, Qijia ^{[3
]}

Zhang, Shanshan ^{[3
]}

Gong, Zhaojian ^{[4
]}

Guo, Can ^{[1
,2
]}

Wang, Fuyan ^{[1
,2
]}

Zeng, Zhaoyang ^{[1
,2
,5
]}

Li, Xiaoling ^{[1
,2
,5
]}

Li, Guiyuan ^{[1
,2
,5
]}

Xiong, Wei ^{[1
,2
,5
]}

Wu, Xu ^{[1
,2
,5
]}

机构：

[1] NHC Key Laboratory of Carcinogenesis and Human Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha,Hunan, China

[2] Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and School of Basic Medicine Sciences, Central South University, Changsha,Hunan, China

[3] Department of Stomatology, Xiangya Hospital, Central South University, Changsha,Hunan, China

[4] Department of Oral and Maxillofacial Surgery, The Second Xiangya Hospital, Central South University, Changsha,Hunan, China

[5] Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Disease Genome Research Center, The Third Xiangya Hospital, Central South University, Changsha,Hunan, China

[6] Department of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha,Hunan, China

来源：

Analytica Chimica Acta | 2022年 / 1189卷

基金：

中国国家自然科学基金;

关键词：

Cell-based - Circbart2.2 - Circular ribonucleic acid - Enzymatic amplifications - Enzyme-assisted - Enzyme-assisted cycling amplification - Exonuclease - Fluorescent sensors - Molecular beacon - Tumour cells;

D O I：

暂无

中图分类号：

学科分类号：

摘要：

Circular Ribonucleic Acid (CircRNA) plays regulatory roles in many biological processes, such as tumors and metabolic diseases. Due to the fact that circRNA is more stable and conservative than linear RNA, circRNA has become a potential biomarker in early clinical diagnosis and biomedical research. Therefore, the quantification of circRNA expression level is of importance for understanding their functions and their applications for disease diagnosis and treatment. Nevertheless, due to the low abundance of circRNA, it is still a challenge for the analysis of circRNA in cells. Herein, we proposed a sensitive detection method for circRNA based on the T7 exonuclease-assisted cycling enzymatic amplification. The fluorescent sensor was constructed by a hairpin molecular beacon and T7 exonuclease. With the cycling enzymatic amplification process, this sensor achieved the limit of detection of 1 pM with a good linear correlation in the range of 0–100 pM (R2 = 0.9891) using circBART2.2 as a model. Furthermore, we applied the proposed method in the determination of circBART2.2 in cell lysates. The results demonstrated that this method has promising applications in early diagnosis of Epstein-Barr virus (EBV) infection-related diseases using circRNA as the biomarker. © 2021 Elsevier B.V.

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