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[小鼠全胚胎RNA原位杂交:一种分析基因表达的有效技术]

[Mouse whole mount RNA in situ hybridization: an effective technique for analyzing gene expression].

作者信息

Xiao Cui-ying, Levanon Ditsa, Zhang Si-zhong, Groner Yoram

机构信息

Department of Medical Genetics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China.

出版信息

Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2004 Feb;21(1):47-51.

Abstract

OBJECTIVE

To set up a method of analyzing gene expression profile from mouse whole embryos.

METHODS

Mouse whole mount RNA in situ hybridization(WM-ISH) of E10.5-E14 embryos was carried out by using digoxigenin-labeled Runx1 and Runx3 RNA probes and their expression profile was observed by detecting the existence and status of corresponding mRNAs in the embryonic tissues.

RESULTS

Clear hybridization signals were observed in different tissues and organs hybridized by Runx1 or Runx3 RNA probe. Different probes and ages of embryos had need of their own optimal proteinase K treatment conditions.

CONCLUSION

Mouse whole mount RNA in situ hybridization is an effective method of analyzing gene expression. It is useful for revealing whole gene expression profile and has a great potentiality in the era of functional genomics. It provides an alternative method of studies on gene expression which is at least as good as LacZ staining and immunohistochemistry. The key factor of the success to mouse whole mount RNA in situ hybridization is whether the proteinase K treatment conditions are optimal or not.

摘要

目的

建立一种分析小鼠全胚胎基因表达谱的方法。

方法

采用地高辛标记的Runx1和Runx3 RNA探针,对E10.5 - E14胚胎进行小鼠全胚胎RNA原位杂交(WM - ISH),通过检测胚胎组织中相应mRNA的存在及状态来观察其表达谱。

结果

用Runx1或Runx3 RNA探针杂交的不同组织和器官中观察到清晰的杂交信号。不同探针及胚胎年龄需要各自最佳的蛋白酶K处理条件。

结论

小鼠全胚胎RNA原位杂交是分析基因表达的有效方法。它有助于揭示全基因表达谱,在功能基因组学时代具有很大潜力。它提供了一种至少与LacZ染色和免疫组织化学一样好的基因表达研究替代方法。小鼠全胚胎RNA原位杂交成功的关键因素是蛋白酶K处理条件是否最佳。

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