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pGreenFire™ Pathway Reporters

Lenti-based monitors of transcription activation

Dual Reporter Lentivector Systems

  • Track postive cells with GFP
  • Quantify level of transactivation with Luciferase
  • Ready-to-transduce virus

Lentiviral Transcriptional Reporter Systems

Eukaryotic gene expression is regulated by a wide variety of developmental and environmental stimuli. First, an extracellular signaling molecule binds to a specific receptor. The signal is then transmitted through a series of molecular cascades, which activate or deactivate specific transcription factors (TFs) that regulate gene expression. The expression of any given gene is controlled by multiple transcription factors, which in turn are modulated by multiple signal transduction pathways. Many of these signal transduction pathways converge at transcription factors that bind to specific transcriptional response elements (TREs) found in the promoters of various genes and modulate the transcription of these genes. The activation of a signal transduction pathway (e.g. by growth factors, drugs, etc.) can therefore be monitored by the expression level of the reporter gene controlled by a promoter containing these response elements.

pGreenFire information

• pGreenFire details
View the pGreenFire lentivector map and features of the reporter system.

• pGreenFire reporter sample data
View pGreenFire reporter sample transactivation data.

Commonly used plasmid-based transcriptional reporter vectors can often skew transcriptional network reporting due to their episomal nature. SBI's lentivector-based transcription reporters integrate into the host's genome and enable proper chromatinization to produce more faithful transcriptional activity reporting.

 

Advantages of lentivector technology include:

  • Ready-to-use pre-packaged constructs with a wide range of Transcriptional Response Elements (TREs) for multiple transcriptional factors.
  • Lentiviral reporter constructs can efficiently transduce nearly all cell types, even those that are difficult-to-transfect such as primary or non-dividing mammalian cells.
  • Our lentiviral-based reporter system is a novel approach to study transcriptional regulation and offers many advantages over current transcription reporter systems. TR constructs will integrate into the genome and therefore be subject to chromatin regulation (Leung, T.H., et.al., 2004). Expression of the reporter gene indicates activation of a given transcriptional response element (TRE) by the cognate transcriptional factor in the natural chromosomal environment rather than in the episomal state in the nucleoplasm as is the case for conventional plasmid-based TR vectors. Tandem copies of integration can be avoided, thus allowing for faithful promoter regulation. Copy number of reporter constructs can be controlled by varying the multiplicity of infection (MOI).
  • Construction of stable reporter cell lines is possible with TR lentivectors in just several days without the need for conventional, low efficiency selection of stable transfectants.
  • Monitoring of signaling pathways by flow cytometry (FACS) is enabled by GFP reporters or quantitate transactivation with Luciferase.