Analysis of messenger RNA expression by in situ hybridization using RNA probes synthesized via in vitro transcription
Analysis of messenger RNA expression by in situ hybridization using RNA probes synthesized via in vitro transcription
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DOI：
10.1016/j.ymeth.2010.08.001
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发表时间：
2010-12-01
期刊：
影响因子：
4.8
通讯作者：
Thompson, Robert C.
中科院分区：
文献类型：
--
作者：
Carter, Bradley S.;Fletcher, Jonathan S.;Thompson, Robert C.
mRNA表达的空间模式的分析对于指定给定基因产物的功能和生理意义至关重要。鉴于哺乳动物基因组中有数万种mRNA，理想情况下，对单个基因功能的全面评估将覆盖表达每种mRNA的特定细胞类型的知识。原位杂交方法代表了一种分子生物学/组织学方法，可以揭示mRNA表达的细胞模式。在这里，我们提出了详细的程序检测特定的mRNA使用放射性RNA探针在组织切片，然后放射自显影检测。这些方法允许对mRNA表达的空间模式进行特异性和灵敏性检测，从而将mRNA表达与细胞类型和功能联系起来。放射性检测方法也有利于mRNA基因表达变化的半定量分析。(C)2010年爱思唯尔公司All rights reserved.
The analysis of the spatial patterning of mRNA expression is critically important for assigning functional and physiological significance to a given gene product. Given the tens of thousands of mRNAs in the mammalian genome, a full assessment of individual gene functions would ideally be overlaid upon knowledge of the specific cell types expressing each mRNA. In situ hybridization approaches represent a molecular biological/histological method that can reveal cellular patterns of mRNA expression. Here, we present detailed procedures for the detection of specific mRNAs using radioactive RNA probes in tissue sections followed by autoradiographic detection. These methods allow for the specific and sensitive detection of spatial patterns of mRNA expression, thereby linking mRNA expression with cell type and function. Radioactive detection methods also facilitate semi-quantitative analyses of changes in mRNA gene expression. (C) 2010 Elsevier Inc. All rights reserved.