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Citations Search

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Nucl. Acids Res. 46. DNA-binding landscape of IRF3, IRF5 and IRF7 dimers: implications for dimer-specific gene regulation. 2018

Andrilenas, K.K., Ramlall, V., Kurland, J., Leung, B., Harbaugh, A.G., Siggers, T.

Notes: Common and dimer-specific DNA binding sites were determined for interferon regulatory factors (IRF3, IRF5, and IRF7). Only IRF3 and IRF7 bound to virus-response elements (VRE) of interferon regulated genes. Mutational analysis of IRFs showed a single residue affected specificity for VRE binding, which was absent in IRF5. The Nano-Glo Dual Luciferase Reporter was used to determine transcriptional activation of IRF variants. (5125)

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114, E2709–8. Hormone and receptor interplay in the regulation of mosquito lipid metabolism. 2017

Wang, X., Hou, Y., Saha, T.T., Pei, G., Raikhel, A.S., and Zou, Z.

Notes: Six mosquitos per sample were frozen in liquid nitrogen and ground in a basic pH lysis buffer then heated to remove NAD+ then neutralized before measuring NADH with the NAD/NADH-Glo™ Assay System. To examine the effect of HNF4 on VLCAD and 3KCT gene expression, the authors employed gel shift assays and luciferase assays of the promoters in pGL4.10[luc2] and control with pGL4.73[hRluc/SV40] transfected into Drosophila S2 cells. Luciferase activities were measured with the Dual-Luciferase® Reporter Assay System on a GloMax® Instrument.  (4834)

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Nucl. Acids Res. 44, 2348-2361. A short conserved motif in ALYREF directs cap- and EJC-dependent assembly of export complexes on spliced mRNAs 2016

Gromadzka, A.M., Steckelberg, A.-L., Singh, K.K, Hofmann, K. and Gehring, N.H.

Notes: NanoLuc® and Firefly luciferase reporters (pCMV128-NanoLuc and pCI-FireflyLuc) were co-transfected into HeLa cells to provide complementary measures of specific mRNA splicing and export activity. In the presence of splicing activity, firefly luciferase pre-mRNA is spliced in the nucleus, exported and translated resulting in firefly luciferase activity. In contrast, the open reading frame of the Nanoluc luciferase in pCMV128 NanoLuc® was placed between two weak splice sites. If the pre-mRNA is spliced, the NanoLuc coding sequence is removed and no luciferase can be expressed. The NanoLuc® reporter can only be translated when the unspliced pre-mRNA is transported to the cytoplasm. pCMV128 NanoLuc® was generated by inserting the NanoLuc® sequence into the NotI and BamHI sites of pCMV128, and then used for export assays. Cells were harvested 72 h post-transfection using 300 μl Passive Lysis Buffer. Lysates were analyzed using the NanoGlo® Dual-Luciferase® Reporter Assay. (4744)

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Nat. Commun. 7, 11624. Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription 2016

Kobayashi, E.H., Suzuki, T., Funayama, R., Nagashima, T., Hayashi, M., Sekine, H., Tanaka, N., Moriguchi, T., Motohashi, H., Nakayama, K. and Yamamoto, M.

Notes: A NanoLuc® luciferase reporter was used to further understand the mechanism of Nrf2 inhibition of proinflammatory cytokine gene induction. Approximately 1 kb upstream region of the IL6 gene TSS containing the WT ARE (GCTGAGTCA) or mutated ARE (AGATCTGAC) was conjugated to the translation start site of the NanoLuc® gene in the pNL2.2 vector. 293T cells (2.0 × 103 cells per well) were plated in 96-well plates 24 h before transfection. The NanoLuc® reporter vectors were co-transfected with pQC-FLAG-hNRF2T80R (constitutively active NRF2) plasmid33 and pcDNA3-p65 plasmid. After 48 hours of the transfection, the luminescence was quantified and normalized using NanoGlo® Dual-Luciferase® Reporter Assay. (4752)

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Cancer Biol Ther 17, 1197–205. Dephosphorylation of the retinoblastoma protein (Rb) inhibits cancer cell EMT via Zeb 2016

Egger, J.V., Lane, M.V., Antonucci, L.A., Dedi, B., and Krucher, N.A. 

Notes: The CellTiter-Glo® 3D Cell Viability Assay was used to measure viability of MDA-MB-468 cells grown on Matrigel with inducible knockdown of PNUTS. The effect of PNUTS knockdown was also assessed in a reporter model of E-Cadherin expression using a firefly luciferase reporter vector and Renilla luciferase control vector. Luciferase activities were measured with the Dual-Luciferase® Reporter Assay System. (4823)

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ACS Chemical Biology 10, 1188–97. Chemogenomic Profiling of Endogenous PARK2 Expression Using a Genome-Edited Coincidence Reporter. 2015

Hasson, S.A., Fogel, A.I., Wang, C., MacArthur, R., Guha, R., Heman-Ackah, S., Martin, S., Youle, R.J,, and Inglese, J.

Notes: These authors describe use of firefly and NanoLuc® luciferases in a coincidence reporter system to screen compounds for their ability to activate transcription of the Parkin gene (PARK2), a potential target for treatment of Parkinson's disease. In a coincidence reporter system, a single transcript containing two luciferase genes is generated. Inclusion of a 2A “ribosomal skipping” sequence between the two luciferase genes enables translation of two reporter proteins. This approach is useful in drug screening because the use of two independent reporters to monitor a single transcriptional event provides a way to distinguish true “hits” from experimental artifacts caused by interaction between library compounds and reporter proteins. True “hits” (compounds that affect transcription of the gene being monitored) cause a signal from both reporters; false positives caused by interaction of compounds with the reporter protein cause a signal from only one reporter. This paper describes the first use of firefly and NanoLuc® luciferases, and the Nano-Glo® Dual-Luciferase® Reporter Assay, in a coincidence reporter system to screen a compound library in an HTS assay. (4565)

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Mol. Endocrinol. 29, 1037-1054. miR-22 and miR-29a Are Members of the Androgen Receptor Cistrome Modulating LAMC1 and Mcl-1 in Prostate Cancer 2015

Pasqualini, L., Bu, H., Puhr, M., Narisu, N., Rainer, J., Schlick, B., Schäfer, G., Angelova, M., Trajanoski, Z., Börno, S.T., Schweiger, M.R., Fuchsberger, C. and Klocker, H.

Notes: The authors investigated the interaction between androgen receptors and miRNAs in order to better understand the role of miRNAs in prostate cancer biology. A NanoLuc® and firefly luciferase miRNA target expression vector (pmirNanoGlo) was used to evaluate the predicted role of miR-22 and miR-29a in the regulation of target genes LAMC1 and MCL1. This bicistronic vector contains NanoLuc® luciferase (NlucP) as the primary reporter gene and Firefly luciferase (Luc2) as the control reporter for normalization. For each target gene, the cDNA fragment predicted to encompass miR-22 and miR-29a target sites, was inserted into multiple cloning regions located in the 3′-UTR of the NanoLuc® reporter vector. PC3 cells were transfected with the reporter vectors in combination with either miR-22 mimic, miR-29a mimic and the Negative Control 4 or antimiR-22, antimiR-29a, and the Negative Control. Transfected PC3 cells underwent NanoLuc® and Firefly luciferase activity measurements using the NanoGlo® Dual-Luciferase® Assay. (4753)

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