Combined microRNA in situ hybridization and immunohistochemical detection of protein markers.
Combined microRNA in situ hybridization and immunohistochemical detection of protein markers.
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DOI：
10.1007/978-1-62703-311-4_22
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Holmstrom, Kim
中科院分区：
文献类型：
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作者：
Nielsen, Boye Schnack;Holmstrom, Kim
MicroRNA是短的（18 - 23个核苷酸）非编码RNA，其通过与mRNA的3'UTR特异性结合而参与基因表达的转录后调节。可以使用特定的锁核酸（LNA）增强探针检测组织中的微小RNA。在microRNA生物标志物发现阶段之后，通过原位检测表征组织中的microRNA表达通常是至关重要的，以便验证候选microRNA生物标志物并允许更好地解释其分子功能和衍生的细胞相互作用。原位杂交数据提供了有关microRNA的背景分布和细胞起源的信息。通过将microRNA原位杂交与蛋白质标记物的免疫组织化学染色相结合，可以精确地表征表达microRNA的细胞并鉴定潜在的microRNA靶标。这种组合技术还可以帮助监测治疗环境中潜在microRNA靶标水平的变化。在本章中，我们提出了一种基于荧光的技术，允许microRNA原位杂交与免疫组织化学的结合，例如miR-21和miR-205与PDCD4和平滑肌α-肌动蛋白的结合的原位检测。
MicroRNAs are short (18-23 nucleotides) non-coding RNAs involved in posttranscriptional regulation of gene expression through their specific binding to the 3'UTR of mRNAs. MicroRNAs can be detected in tissues using specific locked nucleic acid (LNA)-enhanced probes. The characterization of microRNA expression in tissues by in situ detection is often crucial following a microRNA biomarker discovery phase in order to validate the candidate microRNA biomarker and allow better interpretation of its molecular functions and derived cellular interactions. The in situ hybridization data provides information about contextual distribution and cellular origin of the microRNA. By combining microRNA in situ hybridization with immunohistochemical staining of protein markers, it is possible to precisely characterize the microRNA expressing cells and to identify the potential microRNA targets. This combined technology can also help to monitor changes in the level of potential microRNA targets in a therapeutic setting. In this chapter we present a fluorescence-based technology that allows the combination of microRNA in situ hybridization with immunohistochemistry exemplified by the in situ detection of miR-21 and miR-205 in combination with PDCD4 and smooth muscle a-actin.