Reporter gene assay luciferase

Legler J, van den Brink C, Brouwer A, et al. 1998. Assessment of (anti-)estrogenic compounds using a stably transfected luciferase reporter gene assay in the human T47-D breast cancer cell line. Organohalogen Compounds 37 265. 

LEGT.ER J, VAN DEN BRINK C E, BROUWER A, MURK A J, VAN DER SAAG P T, vethaak A D and van der BURG B, Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line , Toxicol Sci 1999 48 55-66. 

George, S.E., P.J. Bungay, and F.H. Naylor. 1997. Evaluation of a CRE-directed luciferase reporter gene assay as an alternative to measuring cAMP accumulation. J. Biomol. Screen. 2, 235-240. 

Figure 2.9. Luciferase reporter gene assay/promoter deletion analysis. Cells are transfected with the CYP1A1 promoter reporter (luciferase) constructs in which the promoter region is increasing deleted. The cells are then treated with benzo[a]pyrene (an inducer of CYP1A1), and cells are harvested 24-48hr later and luciferase activity is measured. Promoter deletion analysis revealed enhancer element between -1140 and -1029. Contained within this region is xenobiotic response element (XRE) to which the aryl hydrocarbon receptor (AHR) binds.

Wfe have recently carried out experiments with cell-based and animal models of inflammatory diseases. The experiments have revealed that human polymorphonuclear cells once activated with phorbol esters to cause an inflammatory response produce copious amounts of chlorinated polyphenol species. Rats treated with lipopolysaccharide (LPS) in an in vivo model of inflammatory disease heavily nitrate polyphenols in tissues (e.g., lung and liver) where inflammatory cell invasion occurs. These modified polyphenols are better antioxidants than their parent compound. Using a luciferase reporter gene assay in COS cells, both chloro- and nitrogenistein were shown to have 1-2 orders lower estrogen receptor activation than genistein itself. In summary, metabolism of polyphenols is rampant, but not always inactivating. 

Figure 3 Luciferase Reporter Gene Assay of Estrogens Bearing Carboranes. COS-1 cells were transfected with EREx5-pGL-TKandpCI-rER and incubated with the compounds at the indicated concentrations (10 IOM to Results are shown as...

Data from a luciferase reporter gene assay using an adherent cell line and comparing 96-and 384-well microplates was kindly supplied by Dr. Eric Gjerstad of Roche Bioscience, Palo Alto, California. 

Table 1. Miniaturization of a Luciferase Reporter Gene Assay. Induced cps (counts per second) refers to the luminescence produced when the reporter gene is activated by a known activator. Control cps refers to the background expression when no activator is present. The S/N is the signal to noise ratio (induced cps/control cps).

Figure 8 (a) Click glycopolymer synthesis and characterization data where x= 1 (Tri), 2 (Tr2), 3 (Tr3), or4 (Tr4).The degree of polymerization has been systematically varied from 35 to 100 [4,26], (b) Transfection data as determined by luciferase reporter gene assays in HeLa cells with polyplexes formed from Tr4 at N/P = 7 (PEI N/P = 5). As the degree of polymerization increases, the transfection efficiency increases, both in the presence and absence of serum (DMEM and Opti-MEM, respectively). In the absence of serum, the toxicity also dramatically increases with the polymer length. Part b adapted from Srinivasachari, S. Liu, Y. Prevette, L. E. Reineke, T. M. Biomaterials 2007, 28,2885. ... 

The estrogenic activities of the synthesized compounds were examined by the luciferase reporter gene assay in which a rat ERK expression plasmid and a reporter plasmid, containing five copies of estrogen-response elanents, were transiently transfected into COS-1 cells. 17P-Estradiol at 1 X 10 -1 X 10" M induced the expression of luciferase in a dose-dependent manner. Among the... 

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