Investigate the activity and interactions of key signal transduction pathways with our Dominant-Negative Vector Sets. Each vector sets includes a vector that expresses a strong wild-type protein and one that provides a dominant negative, nonfunctional version—allowing you to either activate or “knock down” the pathway. To set up a complete assay system, you can co-express these vectors with an appropriate cis-acting reporter vector to measure the activation of a signal transduction pathway and quantify differences in activation.
The p53 Dominant-Negative Vector Set lets you study p53 and related signaling pathways. These vectors constitutively express high levels of wild-type p53 or its dominant-negative mutant. To determine the effects of p53 expression on a particular cell line, simply transfect with the vector containing the gene for wild-type p53, and examine morphology or look for changes in the expression of other pathway proteins.
The IkappaBalpha Dominant-Negative Vector Set provides a convenient way to examine NFkappaB regulation by manipulating its inhibitor, IkappaBalpha, which normally keeps NFkappaB inactive and sequestered in the cytosol. This vector set gives you the ability to compare the effects of overexpressed IkappaBalpha (phosphorylated specifically by your agent) to those of IkappaBalphaM (unable to be phosphorylated under any conditions). By combining these vectors with cis-acting NFkappaB Vectors, you have a complete assay system to study and measure activation of the NFkappaB pathway.
The CREB Dominant-Negative Vector Set includes a constitutively active mutant and two different dominant-negative mutants which allow you to block CREB signaling by separate mechanisms in order to determine which genes are activated by CREB (cAMP response element-binding protein)—a transcription factor that acts downstream in the protein kinase A (PKA) signaling pathway.
Ras & Raf
The Ras Dominant-Negative Vector Set lets you study Ras, a GTP-binding protein that relays signals affecting many cellular processes, including human tumor development (1). We also offer a vector set for studying Raf, the most highly characterized downstream effector of Ras. You can combine the Ras or Raf Vector Sets with our Pathway Profiling Systems to easily monitor cross-talk between different signaling pathways.