دورية أكاديمية

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

التفاصيل البيبلوغرافية
العنوان: A fluorescence strategy for circRNA quantification in tumor cells based on T7 nuclease-assisted cycling enzymatic amplification.
المؤلفون: Qu, Hongke1,2 (AUTHOR), Chen, Mingjian2 (AUTHOR), Ge, Junshang2 (AUTHOR), Zhang, Xiangyan3 (AUTHOR), He, Shuyi2 (AUTHOR), Xiong, Fang4 (AUTHOR), Yan, Qijia4 (AUTHOR), Zhang, Shanshan4 (AUTHOR), Gong, Zhaojian5 (AUTHOR), Guo, Can1,2 (AUTHOR), Wang, Fuyan1,2 (AUTHOR), Zeng, Zhaoyang1,2,6 (AUTHOR), Li, Xiaoling1,2,6 (AUTHOR), Li, Guiyuan1,2,6 (AUTHOR), Xiong, Wei1,2,6 (AUTHOR) xiongwei@csu.edu.cn, Wu, Xu1,2,6 (AUTHOR) wuxu1028@csu.edu.cn
المصدر: Analytica Chimica Acta. Jan2022, Vol. 1189, pN.PAG-N.PAG. 1p.
مصطلحات موضوعية: *CIRCULAR RNA, *EXONUCLEASES, *RNA, *FLUORESCENCE, *EPSTEIN-Barr virus, *CELL determination
مستخلص: 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. [Display omitted] • The circRNA encoded by EBV was specifically detected by fluorescence sensor for the first time. • The enzyme cycle signal amplification strategy greatly improves the sensitivity of detection. • The applicability of this method in real samples was verified by a variety of cell lines. • The developed sensor can be expanded for other circRNA analysis in different diseases. [ABSTRACT FROM AUTHOR]
قاعدة البيانات: Academic Search Index
الوصف
تدمد:00032670
DOI:10.1016/j.aca.2021.339210