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利用 DNA-银纳米簇检测人乳腺癌 MCF-7 细胞中 BRCA1 基因的大片段缺失。

Detection of large deletion in human BRCA1 gene in human breast carcinoma MCF-7 cells by using DNA-Silver Nanoclusters.

机构信息

Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran.

出版信息

Methods Appl Fluoresc. 2017 Dec 13;6(1):015001. doi: 10.1088/2050-6120/aa8988.

Abstract

Here we describe a label-free detection strategy for large deletion mutation in breast cancer (BC) related gene BRCA1 based on a DNA-silver nanocluster (NC) fluorescence upon recognition-induced hybridization. The specific hybridization of DNA templated silver NCs fluorescent probe to target DNAs can act as effective templates for enhancement of AgNCs fluorescence, which can be used to distinguish the deletion of BRCA1 due to different fluorescence intensities. Under the optimal conditions, the fluorescence intensity of the DNA-AgNCs at emission peaks around 440 nm (upon excitation at 350 nm) increased with the increasing deletion type within a dynamic range from 1.0 × 10 to 2.4 × 10 M with a detection limit (LOD) of 6.4 × 10 M. In this sensing system, the normal type shows no significant fluorescence; on the other hand, the deletion type emits higher fluorescence than normal type. Using this nanobiosensor, we successfully determined mutation using the non-amplified genomic DNAs that were isolated from the BC cell line.

摘要

在这里,我们描述了一种基于 DNA-银纳米簇(NC)荧光的无标记检测策略,用于检测乳腺癌(BC)相关基因 BRCA1 中的大片段缺失突变。DNA 模板化银 NCs 荧光探针与靶 DNA 的特异性杂交可以作为增强 AgNCs 荧光的有效模板,可用于区分由于荧光强度不同而导致的 BRCA1 缺失。在最佳条件下,在发射峰处约为 440nm(在 350nm 激发下)的 DNA-AgNCs 的荧光强度随着从 1.0×10 到 2.4×10 M 的动态范围内的缺失类型的增加而增加,检测限(LOD)为 6.4×10 M。在这个传感系统中,正常类型没有明显的荧光;另一方面,缺失类型的荧光发射比正常类型更高。使用这种纳米生物传感器,我们成功地使用从 BC 细胞系分离的非扩增基因组 DNA 来确定突变。

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