基于G-四链体模板的银纳米簇共振能量转移比率检测miRNA

发布时间：2018-04-28 03:02

本文选题：G-四链体 + 银纳米簇　；参考：《化学学报》2017年11期

【摘要】：以DNA为模板合成的荧光银纳米簇(DNA-Ag NC)是一类性能优秀的发光探针,已被广泛应用于传感与成像领域.目前大多数定量方法都是基于目标物结合带来的直接的银纳米簇发光强度的变化,因此,分析方法的重现性将不可避免地受实验场地或测定条件变化的影响.本工作中,采用以G-四链体为模板的银纳米簇(GQ-AgNC)与菁染料Cyanine5(Cy5)为能量供-受体对,发展了基于荧光共振能量转移(FRET)机理的mi RNA比率测定方法.其中,Cy5标签具有通用性,简化了实际应用中的实验设计,降低了试剂成本.针对mi RNA let-7a的方法,检测动态范围为12~300nmol/L,检测限为6.9 nmol/L,且可以将let-7a与let-7家族中的其他mi RNA区分.HepG2细胞的总RNA提取物加标回收结果表明该方法具有应用于临床样本测试的潜力.本研究工作拓展了DNA-Ag NC的应用,并有助于加深在FRET设计中以DNA-Ag NC为供体进行分析应用的进一步理解.
[Abstract]:The fluorescent silver nanoclusters DNA-Ag NCs synthesized by using DNA as template are a kind of excellent luminescent probes and have been widely used in sensing and imaging fields. At present, most quantitative methods are based on the direct changes in luminescence intensity of silver nanoclusters caused by the combination of target materials. Therefore, the reproducibility of analytical methods will inevitably be affected by the changes of experimental sites or determination conditions. In this work, the GQ-AgNC-based silver nanoclusters (GQ-AgNC) and cyanine dye Cyanine (Cyanine 5C5) were used as energy donor / acceptor pairs to develop a method for the determination of mi RNA ratio based on the fluorescence resonance energy transfer (FRET) mechanism. The tag of Cy5 is universal, which simplifies the experimental design in practical application and reduces the cost of reagents. For the method of mi RNA let-7a, the dynamic range of detection was 123nmol / L, the detection limit was 6.9 nmol / L, and the total RNA extract of let-7a differentiated from other mi RNA in the let-7 family could be labeled and recovered. The results showed that the method had the potential to be applied to clinical sample testing. This work extends the application of DNA-Ag NC and helps to further understand the application of DNA-Ag NC in FRET design.
【作者单位】：北京分子科学国家实验室生物有机与分子工程教育部重点实验室北京大学化学与分子工程学院分析化学所;
【基金】：国家自然科学基金(Nos.21475004和21535006)资助~~
【分类号】：O629.7;O657.3

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