The main role of the cAMP response element, or CRE, is mediating the effects of Protein Kinase A (PKA) by way of transcription. Upon phosphorylation, CREB forms a functionally active dimer that binds the CRE element within the promoters of target genes and activates transcription. CRE is at the focus of many extracellular and intracellular signaling pathways, including cAMP, calcium, GPCR (G-protein coupled receptors) and neurotrophins. The cAMP/PKA signaling pathway is critical to numerous life processes in living organisms.

The CRE/CREB Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by multimerized cAMP response element (CRE) located upstream of the minimal TATA promoter (Figure 1). After transduction, activation of the cAMP/PKA signaling pathway in the target cells can be monitored by measuring the luciferase activity. 

The lentiviruses were produced from HEK293T cells in medium containing 90% DMEM + 10% FBS.

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Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Screen for activators or inhibitors of cAMP/PKA signaling pathway in the transduced target cells.

1. Pessara, U., Koch, N. (1990) Tumor necrosis factor alpha regulates expression of the major histocompatibility complex class II-associated invariant chain by binding of an NF-κB-like factor to a promoter element. Mol Cell Biol. 10(8):4146-4154.

2. Baeuerle, P.A. (1998) Pro-inflammatory signaling: last pieces in the NF-κB puzzle? Curr Biol. 8(1):R19-R22.

The lentiviruses are produced with the third generation SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal.

Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Cell Signaling Pathway

Cell Line