1.Mouse whole mount RNA in situ hybridization: an effective technique for analyzing gene expression.
Cui-ying XIAO ; Ditsa LEVANON ; Si-zhong ZHANG ; Yoram GRONER
Chinese Journal of Medical Genetics 2004;21(1):47-51
OBJECTIVETo set up a method of analyzing gene expression profile from mouse whole embryos.
METHODSMouse 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.
RESULTSClear 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.
CONCLUSIONMouse 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.
Animals ; Core Binding Factor Alpha 2 Subunit ; Core Binding Factor Alpha 3 Subunit ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; metabolism ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Developmental ; In Situ Hybridization ; methods ; Mice ; Proto-Oncogene Proteins ; genetics ; RNA ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Sensitivity and Specificity ; Transcription Factors ; genetics