( New tools to study in vivo functions ...)In Vivo Signal Transduction Pathway Reporting Systems

New tools to study in vivo functions of new genes and drug candidates

Li Xu * Tim Sanchez * ChaoFeng Zheng
Stratagene Cloning Systems, Inc.

Stratagene's PathDetect in vivo signal transduction pathway reporting systems are designed for specific, rapid and convenient assessment of the in vivo activation of signal transduction pathways. These reporting systems can be used to study the in vivo effects of new genes, growth factors and drug candidates on the activation of c-Jun N-terminal kinase (JNK), mitogen-activated protein kinase (MAPK), cyclic AMP-dependent protein (PKA) and other signaling molecules leading to the activation of these kinases. When a fusion activator plasmid, reporter plasmid and uncharacterized gene are cotransfected into mammalian cells, either direct or indirect phosphorylation of the fusion activator protein by the uncharacterized gene product will activate transcription of the luciferase gene from the reporter plasmid. Similarly, the effects of extracellular stimuli can be studied with these systems. Very low background and as high as 200fold activation have been observed with the PathDetect c-Jun, Elk1 and CREB transreporting systems.

Signal transduction is essential for cellular growth, differentiation and the coordination of various cellular activities in multicellular organisms. Many human diseases are caused by disturbances in signal transduction. Through standard techniques in molecular biology, hundreds of genes encoding receptors, protein kinases, phosphatases and other regulatory proteins have been cloned, and many of these gene products are implicated in signal transduction pathways. Because most of the standard cloning methods do not yield information about the function of gene products, other methods are needed in order to assess the in vivo function or activity of such proteins.

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