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一种基于银纳米簇和与化学输入信号响应的 NAND 逻辑门的共振瑞利散射传感器,用于灵敏地区分端粒 DNA 长度和监测特殊基序(G-四链体和 i- 发夹)。

A resonance Rayleigh scattering sensor for sensitive differentiation of telomere DNA length and monitoring special motifs (G-quadruplex and i-motif) based on the Ag nanoclusters and NAND logic gate responding to chemical input signals.

机构信息

The Key Laboratory of Life-Organic Analysis, Qufu Normal University, Qufu, 273165, Shandong, China.

The Key Laboratory for Colloid and Interface Chemistry of Education Ministry, Shandong University, Jinan, 250100, Shandong, China.

出版信息

J Nanobiotechnology. 2018 Oct 9;16(1):78. doi: 10.1186/s12951-018-0407-5.

Abstract

BACKGROUND

Differentiation of telomere length is of vital importance because telomere length is closely related with several deadly diseases such as cancer. Additionally, G-quadruplex and i-motif formation in telomeric DNA have been shown to act as a negative regulator of telomere elongation by telomerase in vivo and are considered as an attractive drug target for cancer chemotherapy.

RESULTS

In this assay, Ag nanoclusters templated by hyperbranched polyethyleneimine (PEI-Ag NCs) are designed as a new novel resonance Rayleigh scattering (RRS) probe for sensitive differentiation of telomere length and monitoring special motifs (G-quadruplex and i-motif). In this assay, free PEI-Ag NC probe or DNA sequence alone emits low intensities of RRS, while the formation of PEI-Ag NCs/DNA complexes yields greatly enhanced RRS signals; however, when PEI-Ag NCs react with G-quadruplex or i-motif, the intensities of RRS exhibit slight changes. At the same concentration, the enhancement of RRS signal is directly proportional to the length of telomere, and the sensitivity of 64 bases is the highest with the linear range of 0.3-50 nM (limit of detection 0.12 nM). On the other hand, due to the conversion of telomere DNA molecules among multiple surrounding conditions, a DNA logic gate is developed on the basis of two chemical input signals (K and H) and a change in RRS intensity as the output signal.

CONCLUSION

Our results indicate that PEI-Ag NCs can serve as a novel RRS probe to identify DNA length and monitor G-quadruplex/i-motif through the different increasing degrees of RRS intensity. Meanwhile, the novel attributes of the nanoprobe stand superior to those involving dyes or labeled DNA because of no chemical modification, low cost, green, and high efficiency.

摘要

背景

端粒长度的分化至关重要,因为端粒长度与癌症等几种致命疾病密切相关。此外,已证明在体内端粒 DNA 中的 G-四链体和 i- 发夹结构的形成可作为端粒酶延长端粒的负调节剂,并被认为是癌症化疗的有吸引力的药物靶标。

结果

在该测定中,超支化聚乙烯亚胺(PEI)模板的银纳米团簇(PEI-Ag NCs)被设计为一种新的新型共振瑞利散射(RRS)探针,用于灵敏地区分端粒长度并监测特殊基序(G-四链体和 i-发夹)。在该测定中,游离的 PEI-Ag NC 探针或 DNA 序列本身发射出低强度的 RRS,而形成 PEI-Ag NCs/DNA 复合物则产生大大增强的 RRS 信号;然而,当 PEI-Ag NCs 与 G-四链体或 i-发夹反应时,RRS 的强度会发生轻微变化。在相同的浓度下,RRS 信号的增强与端粒的长度成正比,并且 64 个碱基的灵敏度最高,线性范围为 0.3-50 nM(检测限为 0.12 nM)。另一方面,由于端粒 DNA 分子在多种环境条件下的转换,基于两个化学输入信号(K 和 H)和 RRS 强度变化作为输出信号,开发了 DNA 逻辑门。

结论

我们的结果表明,PEI-Ag NCs 可以用作新型 RRS 探针,通过不同程度的 RRS 强度增加来识别 DNA 长度并监测 G-四链体/i-发夹。同时,由于无需化学修饰,成本低,绿色且高效,纳米探针的新颖特性优于涉及染料或标记 DNA 的特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/872a/6176526/1064e3abda1a/12951_2018_407_Fig1_HTML.jpg

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