利用邮政编码阵列系统同时检测不同MicroRNA类型。

IF 1.3 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Journal of Nucleic Acids Pub Date : 2013-01-01 Epub Date: 2013-09-02 DOI:10.1155/2013/496425
Sonja U Weishaupt, Steffen Rupp, Karin Lemuth
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引用次数: 6

摘要

MicroRNAs (miRNAs)是基因表达的重要负调控因子。它们在肿瘤发生中的意义是基于它们在许多人类癌症疾病中的失调。有趣的是,在肿瘤细胞中,前体和成熟miRNA水平的比例发生了变化。因此,miRNA类型水平的差异作为生物标志物具有很高的潜力,相对高通量的检测可能允许更准确地表征亚型,特别是在非常异质性的肿瘤实体的情况下。存在几种分子方法用于检测成熟和前体mirna。DNA微阵列被认为是一种高通量的肿瘤miRNA综合检测方法。然而,这两种miRNA类型的同时基于阵列的检测是有限的,因为成熟的miRNA序列在这两种形式中是相同的。在这里,我们提出了一个基于邮政编码DNA微阵列的系统,结合一种新的标记方法,可以在一个单一的实验中同时检测前体和成熟的mirna。使用合成的miRNA模板,我们证明了该方法对不同miRNA类型的特异性,以及高达四个数量级的检测范围。此外,在人肿瘤细胞中检测到成熟mirna和前体mirna并进行了验证。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Simultaneous Detection of Different MicroRNA Types Using the ZIP-Code Array System.

Simultaneous Detection of Different MicroRNA Types Using the ZIP-Code Array System.

Simultaneous Detection of Different MicroRNA Types Using the ZIP-Code Array System.

Simultaneous Detection of Different MicroRNA Types Using the ZIP-Code Array System.

MicroRNAs (miRNAs) are important negative regulators of gene expression. Their implication in tumorigenesis is based on their dysregulation in many human cancer diseases. Interestingly, in tumor cells, an altered ratio of precursor and mature miRNA levels has been described. Consequently, differences in miRNA type levels have a high potential as biomarkers and comparative high-throughput-based detection might permit a more accurate characterization of subtypes, especially in the case of very heterogeneous tumor entities. Several molecular methods exist for the detection of mature and precursor miRNAs. DNA microarrays are predestinated as a high-throughput method for comprehensive miRNA detection in tumors. However, the simultaneous array-based detection of both these miRNA types is limited because the mature miRNA sequence is identically present in both forms. Here we present a ZIP-code DNA microarray-based system in combination with a novel labeling approach, which enables the simultaneous detection of precursor and mature miRNAs in one single experiment. Using synthetic miRNA templates, we demonstrate the specificity of the method for the different miRNA types, as well as the detection range up to four orders of magnitude. Moreover, mature and precursor miRNAs were detected and validated in human tumor cells.

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来源期刊
Journal of Nucleic Acids
Journal of Nucleic Acids BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
3.10
自引率
21.70%
发文量
5
审稿时长
12 weeks
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