GeneBLAzer technology overview

Thermo Fisher´s GeneBLAzer® beta-lactamase reporter technology provides you with a reliable, integrated set of tools for accelerating cell-based target validation, pathway analysis, and compound screening in drug discovery. This proven technology combines molecular and cell biology and a Fluorescence Resonance Energy Transfer (FRET)-based, ratiometric detection method that reduces experimental noise which can lead to false hits. The GeneBLAzer® Technology consists of readily accessible tools and custom assays for studying numerous target classes and cellular processes, including: surface and intracellular reporters (such as nuclear and cytokine receptors, orphan and known G-protein coupled receptors), a wide range of signal transduction pathways, ion channels, other transporters, and transcriptional regulators (including kinases and intracellular processes).

 

GeneBLAzer® Technology uses a mammalian-optimized bla gene combined with a FRET-enabled substrate to provide reliable and sensitive detection in cells. Cells are loaded with an engineered fluorescent substrate containing two fluoroprobes, coumarin and fluorescein. In the absence of bla expression, the substrate molecule remains intact. In this state, excitation of the coumarin results in fluorescence resonance energy transfer to the fluorescein moiety and emission of green light. However, in the presence of bla expression, the substrate is cleaved, separating the fluorophores, and disrupting energy transfer. Excitation of the coumarin in the presence of enzyme bla activity results in a blue fluorescence signal. The resulting blue:green ratio provides a normalized reporter response.

 

Fig. 1: Fluorescent detection of β-lactamase reporter gene response using GeneBLAzer® Technology. After substrate loading, in the absence of β-lactamase expression, cells generate green fluorescence. In the presence of β-lactamase expression, the substrate is cleaved and cells generate blue fluorescence.

For more information about GeneBLAzer® Technology and related cellular assays for different drug target classes available from Life Technologies, please see their website:

 

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