Engineered luciferases for molecular sensing in living cells

Promega Corporation, Madison, WI 53711, USA.
Current Opinion in Biotechnology (Impact Factor: 8.04). 04/2009; 20(1):14-8. DOI: 10.1016/j.copbio.2009.02.013
Source: PubMed

ABSTRACT As a means for visualizing molecular physiology within living cells, new strategies are emerging for engineering luciferases into intracellular biosensors. These biosensors can be classified as bimolecular, relying on complementation of luciferase fragments, or intramolecular, relying on domain insertion within the luciferase structure. Multiple design strategies have recently surfaced for the development of intramolecular sensors, allowing dynamic detection of small molecules or post-translational modifications within cells. Building on successes achieved in cell culture, these sensors are now beginning to reveal molecular processes within living organisms.

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    • "developmental biology (Akiyoshi et al., 2014), medical research (Horibe et al., 2014; Sramek et al., 2011), signal transduction analysis (Hall et al., 2012; Roger et al., 2008; Sugiyama et al., 2014), molecular interaction analysis (Binkowski et al., 2009; Cosby, 2009), and radiation biology (Pratx et al., 2012, 2013). "
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    Microscopy Research and Technique 06/2015; DOI:10.1002/jemt.22529 · 1.17 Impact Factor
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    • "Not only is luciferase activity readily measurable in a broad dynamic range with high sensitivity, luciferase activity can be measured continuously over time in response to various conditions making luciferase an excellent model enzyme to investigate deactivation. In addition to its broad use in gene reporter assays and in various bioanalytical assays (ATP detection down to attomole concentrations are possible), firefly luciferase was also used to study protein refolding as well as the effects of MHz electromagnetic radiation on biological systems [12] [13] [14]. Since luciferase is a fairly " typical " monomeric enzyme, detailed understanding of mechanisms of luciferase thermal stability will advance specific applications and can contribute to general understanding of mechanisms that affect stability of this and other enzymes under various conditions. "
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    Biochimica et Biophysica Acta 10/2011; 1814(10):1318-24. DOI:10.1016/j.bbapap.2011.06.010 · 4.66 Impact Factor
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    • "Genetic manipulation of firefly luciferase into a reversible biosensor of cAMP generation involved the insertion of the cAMP binding domain B from the regulatory subunit type IIb into a circularly permuted form of firefly luciferase (Fan et al., 2008; Binkowski et al., 2009). In vitro, the firefly luciferasebased cAMP biosensor has a pEC50 for cAMP of 6.3 and a large signal-to-noise window of approximately 70-fold (Fan et al., 2008). "
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