Rational design of a novel aptamer-based biosensor for a target enzyme via modification of GFP-like fluorogens: Carboxylesterase 2A as a case study
Rational design of a novel aptamer-based biosensor for a target enzyme via modification of GFP-like fluorogens: Carboxylesterase 2A as a case study
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通过类 GFP 荧光剂的修饰合理设计一种新型基于适体的靶酶生物传感器：以羧酸酯酶 2A 为例
DOI：
10.1016/j.snb.2020.129312
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发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Guang-Bo Ge
中科院分区：
文献类型：
--
作者：
Yue Chen;Qing-Qing He;Dan-Dan Wang;Fang-Yuan Wang;Xiao-Qing Guan;Qing Hu;Hao-Nan Wang;Li-Wei Zou;Qing-Feng Tang;Yi-Nan Wang;Guang-Bo Ge
本文采用合理的设计策略，通过修饰绿色荧光蛋白（GFP）样荧光团作为关键识别元件，构建了一种新型实用的核酸适体生物传感器，用于检测和成像人羧酸酯酶2A（CES 2A）。为此，根据靶酶的底物选择性和RNA适配体-荧光团复合物的荧光检测机理，设计了一系列3，5-二氟-4-羟基苯亚甲基咪唑啉酮（DFHBI）酯类化合物，并通过对接模拟和实验分析评价了其作为CES 2A底物的潜力。结果表明，至少有4种DFHBI酯可被CES 2A水解，其中T-DFHBI（DFHBI三甲基乙酰酯）表现出化学稳定性、快速响应和对CES 2A高度特异性的最佳组合。在信号增强子（Broccoli，一种RNA适体）的辅助下，T-DFHBI在低至8.8ng/mL的水平下测量CES 2A。新开发的基于适体的生物传感器显示出良好的性能，用于感测和成像真实的样品中的CES 2A活性，包括组织制备物和活细胞。总的来说，这项研究表明了一种新的策略，合理的设计和开发的适体为基础的生物传感器的目标酶（S），而这种策略可以扩展到开发其他发光的适体为基础的生物传感器的范围广泛的酶，特别是水解酶。
Herein, a rational design strategy was used to construct a novel and practical aptamer-based biosensor for sensing and imaging human carboxylesterase 2A (CES2A),viamodification of a green fluorescent protein (GFP)-like fluorogen as a key recognition element. For this purpose, series of 3, 5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) esters were designed based on the substrate preference of the target enzyme and the mechanism of RNA aptamer–fluorogen complex for fluorescence detection, while their potentials as CES2A substrates were evaluated by docking simulations and experimental assays. The results demonstrated that at least four DFHBI esters could be readily hydrolyzed by CES2A, whileT-DFHBI(DFHBI trimethylacetyl ester) showed the best combination of chemical stability, fast response and high specificity towards CES2A. Aided by the signal enhancer (Broccoli, a RNA aptamer),T-DFHBImeasured CES2A at the level as low as 8.8 ng/mL. The newly developed aptamer-based biosensor displayed good performance for sensing and imaging of CES2A activities in real samples, including tissue preparations and living cells. Collectively, this study demonstrates a new strategy for rational design and development of aptamer-based biosensors for target enzyme(s), while such strategy can be expanded to develop other light-up aptamer-based biosensors for a wide range of enzymes, particularly for the hydrolases.
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基于 DNA 杂交扩增的核酸和适体特异性配体检测的通用比色法
影响因子：
2.9
作者：
Li, Shuang;Shang, Xinxin;You, Jinmao
通讯作者：
You, Jinmao
DOI：
10.1126/science.1207339
发表时间：
2011-07-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
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通讯作者：
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DOI：
10.1039/c5cc01526j
发表时间：
2015-05-28
期刊：
Chemical communications (Cambridge, England)
影响因子：
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作者：
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通讯作者：
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影响因子：
9.5
作者：
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影响因子：
7.4
作者：
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