适体
化学
DNA
鸟嘌呤
纳米团簇
杂交探针
荧光
序列(生物学)
识别序列
A-DNA
生物物理学
癌细胞
计算生物学
分子生物学
组合化学
生物化学
癌症
核苷酸
基因
生物
遗传学
限制性酶
有机化学
物理
量子力学
作者
Jinjin Yin,Xiaoxiao He,Kemin Wang,Fugui Xu,Jingfang Shangguan,Dinggeng He,Hui Shi
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2013-11-27
卷期号:85 (24): 12011-12019
被引量:171
摘要
We present here a label-free and turn-on aptamer strategy for cancer cell detection based on the recognition-induced conformation alteration of aptamer and hybridization-induced fluorescence enhancement effect of DNA–silver nanoclusters (DNA–Ag NCs) in proximity of guanine-rich DNA sequences. In this strategy, two tailored DNA probes were involved. One is designed as a hairpin-shaped structure consisting of a target specific aptamer sequence at the 3′-end, a guanine-rich DNA sequence, and an arm segment at the 5′-end (denote as recognition probe). The other, serving as a signal probe, contains a sequence for Ag NCs templated synthesis and a link sequence complementary to the arm segment of the recognition probe. Recognizing and binding of the aptamer to cancer cells enforces the recognition probe to undergo a conformational alteration and then initiates hybridization between the arm segment of the recognition probe and the link sequence of the signal probe. The Ag NCs are then close to the guanine-rich DNA, leading to an enhanced fluorescence readout. As proof-of-concept, the CCRF-CEM cancer cell detection were performed by using the specific aptamer, sgc8c. It was demonstrated that this strategy could specially image the CCRF-CEM cells. Determination by flow cytometry allowed for detection of as low as 150 CCRF-CEM cells in 200 μL binding buffer. The general applicability of the strategy is also achieved in the successful detection of Ramos cells. These results implied that this strategy holds considerable potential for simple, sensitive, universal, and specific cancer cell detection with no required washing and separation steps.
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