Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes
Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes
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DOI：
10.1016/j.ymeth.2010.07.002
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发表时间：
2010-12-01
期刊：
影响因子：
4.8
通讯作者：
Nielsen, Boye Schnack
中科院分区：
文献类型：
--
作者：
Jorgensen, Stine;Baker, Adam;Nielsen, Boye Schnack
微小核糖核酸（miRNAs）是一组通常长度为18 - 22个碱基对的小分子非编码核糖核酸分子。miRNAs被认为是转录后水平基因表达的负调控因子，在正常生理发育、组织内稳态和疾病中具有重要作用。单个微小核糖核酸的细胞起源对于理解它们在多细胞和复杂结构组织的分子和生物学过程中的作用至关重要。为了在临床和实验用福尔马林固定、石蜡包埋的样本中对miRNAs进行定位，我们基于使用双地高辛（DIG）标记的锁核酸 - 脱氧核糖核酸（LNA - DNA）嵌合探针，开发了一种简单且稳健的单日原位杂交方案。我们表明该方案能够进行特异性分析，并展示了在人类和小鼠石蜡材料中对miR - 1、miR - 21、miR - 124、miR - 126、miR - 145和miR - 205的检测。这些微小核糖核酸已明确的定位使它们在针对特定组织和微小核糖核酸优化微小核糖核酸原位杂交检测时，成为理想的参照微小核糖核酸。（C）2010年由爱思唯尔公司出版
MicroRNAs (miRNAs) constitute a group of small non-coding RNA molecules generally 18-22 base-pairs in length. miRNAs are considered to be negative regulators of gene expression at the level of post-transcription and are important in normal physiological development, tissue homeostasis and disease. The cellular origin of individual microRNAs is of utmost importance for understanding their roles in molecular and biological processes, in multi-cellular and complex structured tissues. For the localization of miRNAs in clinical and experimental formalin-fixed and paraffin-embedded samples we have developed a simple and robust one-day in situ hybridization protocol based on the use of double digoxigenin (DIG)labeled LNA-DNA chimeric probes. We show that the protocol enables analyses of specificity, and demonstrate the detection of miR-1, miR-21, miR-124, miR-126, miR-145, and miR-205 in human and murine paraffin material. The well established localization of these microRNAs makes them ideal for use as reference microRNAs when optimizing the microRNA in situ hybridization assay for a particular tissue and miRNA. (C) 2010 Published by Elsevier Inc.