Optimization of a cAMP response element signal pathway reporter system

Shan, Qiang and Storm, Daniel R. (2010) Optimization of a cAMP response element signal pathway reporter system. Journal of Neuroscience Methods, 191 1: 21-25. doi:10.1016/j.jneumeth.2010.06.003


Author Shan, Qiang
Storm, Daniel R.
Title Optimization of a cAMP response element signal pathway reporter system
Journal name Journal of Neuroscience Methods   Check publisher's open access policy
ISSN 0165-0270
1872-678X
Publication date 2010-08
Sub-type Article (original research)
DOI 10.1016/j.jneumeth.2010.06.003
Volume 191
Issue 1
Start page 21
End page 25
Total pages 5
Place of publication Amsterdam, Netherlands
Publisher Elsevier BV
Collection year 2011
Language eng
Abstract A sensitive cAMP response element (CRE) reporter system is essential for studying the cAMP/protein kinase A/cAMP response element binding protein signal pathway. Here we have tested a few CRE promoters and found one with high sensitivity to external stimuli. Using this optimal CRE promoter and the enhanced green fluorescent protein as the reporter, we have established a CRE reporter cell line. This cell line can be used to study the signal pathway by fluorescent microscope, fluorescence-activated cell analysis and luciferase assay. This cell line's sensitivity to forskolin, using the technique of fluorescence-activated cell sorting, was increased to approximately seven times that of its parental HEK 293 cell line, which is currently the most commonly used cell line in the field for the signal pathway study. Therefore, this newly created cell line is potentially useful for studying the signal pathway's modulators, which generally have weaker effect than its mediators. Our research has also established a general procedure for optimizing transcription-based reporter cell lines, which might be useful in performing the same task when studying many other transcription-based signal pathways. © 2010 Elsevier B.V.
Keyword cAMP
cAMP response element
Cell signaling pathway
Reporter cell line
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2011 Collection
Queensland Brain Institute Publications
 
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Created: Sun, 05 Sep 2010, 00:04:34 EST