当前位置: X-MOL 学术ACS Sens. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
One-Pot Identification of BCR/ABLp210 Transcript Isoforms Based on Nanocluster Beacon
ACS Sensors ( IF 8.2 ) Pub Date : 2021-07-29 , DOI: 10.1021/acssensors.1c00695
Xiaolong Gou 1 , Lulu Xu 1, 2 , Suqing Yang 3 , Xiaoxue Cheng 1 , Haiping Wu 4 , Decai Zhang 1 , Weicheng Shi 1 , Shijia Ding 4 , Yuhong Zhang 1 , Wei Cheng 1
Affiliation  

The BCR/ABLp210 fusion gene is a classic biomarker of chronic myeloid leukemia, which can be divided into e13a2 and e14a2 isoforms according to different breakpoints. These two isoforms showed distinct differences in clinical manifestation, treatment effect, and prognosis risk. Herein, a strategy based on nanocluster beacon (NCB) fluorescence was developed to identify the e13a2 and e14a2 isoforms in one-pot. Because the fluorescence of AgNCs can be activated when they are placed in proximity to the corresponding enhancer sequences, thymine-rich (T-rich) or guanine-rich (G-rich). In this work, we explored an ideal DNA–AgNCs template as an excellent molecular reporter with a high signal-to-noise ratio. After recognition with the corresponding isoforms, the AgNCs can be pulled closer to the T-rich or G-rich sequences to form a three-way junction structure and generate fluorescence with corresponding wavelengths. Therefore, by distinguishing the corresponding wavelengths of AgNCs, we successfully identified two isoforms in one tube with the limitation of 16 pM for e13a2 and 9 pM for e14a2. Moreover, this strategy also realized isoform identification in leukemia cells and newly diagnosed CML patients within 40 min, which provides a powerful tool to distinguish fusion gene subtypes at the same time.

中文翻译:

基于纳米簇信标的 BCR/ABLp210 转录本异构体的一锅鉴定

BCR/ABL p210融合基因是慢性粒细胞白血病的经典生物标志物,根据不同的断点可分为e13a2和e14a2亚型。这两种异构体在临床表现、治疗效果和预后风险方面表现出明显差异。在此,开发了一种基于纳米簇信标 (NCB) 荧光的策略来识别一锅中的 e13a2 和 e14a2 亚型。因为当 AgNCs 靠近相应的增强子序列、富含胸腺嘧啶 (T-rich) 或富含鸟嘌呤 (G-rich) 时,它们的荧光可以被激活。在这项工作中,我们探索了理想的 DNA-AgNCs 模板作为具有高信噪比的优秀分子报告基因。与相应的异构体识别后,AgNCs 可以被拉近 T-rich 或 G-rich 序列,形成三路连接结构,并产生相应波长的荧光。因此,通过区分 AgNCs 的相应波长,我们成功地在一个管中鉴定了两种同工型,e13a2 的限制为 16 pM,e14a2 的限制为 9 pM。此外,该策略还实现了在40分钟内对白血病细胞和新诊断的CML患者进行亚型鉴定,为同时区分融合基因亚型提供了有力工具。
更新日期:2021-08-27
down
wechat
bug