Reporter Enzyme Inhibitor Study To Aid Assembly of Orthogonal Reporter Gene Assays

2013 ◽  
Vol 8 (5) ◽  
pp. 1009-1017 ◽  
Author(s):  
Pei-i Ho ◽  
Kimberley Yue ◽  
Pramod Pandey ◽  
Lyne Breault ◽  
Fred Harbinski ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Enzyme Inhibitor ◽  
Reporter Enzyme ◽  
Inhibitor Study ◽  
Reporter Gene Assays

Influence of chlorophyll and tannins in plant extracts on cell-based luciferase reporter gene assays

Planta Medica ◽  
2009 ◽  
Vol 75 (09) ◽  
Author(s):  
S Vogl ◽  
P Picker ◽  
N Fakhrudin ◽  
A Atanasov ◽  
E Heiß ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Plant Extracts ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Reporter Gene Assays

scholarly journals GATA-targeted compounds modulate cardiac subtype cell differentiation in dual reporter stem cell line

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Mika J. Välimäki ◽  
Robert S. Leigh ◽  
Sini M. Kinnunen ◽  
Alexander R. March ◽  
Ana Hernández de Sande ◽  
...  
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Cell Fate ◽  
Reporter Gene ◽  
Gene Regulatory Networks ◽  
Regulatory Networks ◽  
Cell Fate Decisions ◽  
Dual Reporter ◽  
Gene Regulatory ◽  
Reporter Gene Assays

AbstractBackgroundPharmacological modulation of cell fate decisions and developmental gene regulatory networks holds promise for the treatment of heart failure. Compounds that target tissue-specific transcription factors could overcome non-specific effects of small molecules and lead to the regeneration of heart muscle following myocardial infarction. Due to cellular heterogeneity in the heart, the activation of gene programs representing specific atrial and ventricular cardiomyocyte subtypes would be highly desirable. Chemical compounds that modulate atrial and ventricular cell fate could be used to improve subtype-specific differentiation of endogenous or exogenously delivered progenitor cells in order to promote cardiac regeneration.MethodsTranscription factor GATA4-targeted compounds that have previously shown in vivo efficacy in cardiac injury models were tested for stage-specific activation of atrial and ventricular reporter genes in differentiating pluripotent stem cells using a dual reporter assay. Chemically induced gene expression changes were characterized by qRT-PCR, global run-on sequencing (GRO-seq) and immunoblotting, and the network of cooperative proteins of GATA4 and NKX2-5 were further explored by the examination of the GATA4 and NKX2-5 interactome by BioID. Reporter gene assays were conducted to examine combinatorial effects of GATA-targeted compounds and bromodomain and extraterminal domain (BET) inhibition on chamber-specific gene expression.ResultsGATA4-targeted compounds 3i-1000 and 3i-1103 were identified as differential modulators of atrial and ventricular gene expression. More detailed structure-function analysis revealed a distinct subclass of GATA4/NKX2-5 inhibitory compounds with an acetyl lysine-like domain that contributed to ventricular cells (%Myl2-eGFP+). Additionally, BioID analysis indicated broad interaction between GATA4 and BET family of proteins, such as BRD4. This indicated the involvement of epigenetic modulators in the regulation of GATA-dependent transcription. In this line, reporter gene assays with combinatorial treatment of 3i-1000 and the BET bromodomain inhibitor (+)-JQ1 demonstrated the cooperative role of GATA4 and BRD4 in the modulation of chamber-specific cardiac gene expression.ConclusionsCollectively, these results indicate the potential for therapeutic alteration of cell fate decisions and pathological gene regulatory networks by GATA4-targeted compounds modulating chamber-specific transcriptional programs in multipotent cardiac progenitor cells and cardiomyocytes. The compound scaffolds described within this study could be used to develop regenerative strategies for myocardial regeneration.

Chemiluminescent and Bioluminescent Reporter Gene Assays

1994 ◽  
Vol 219 (2) ◽  
pp. 169-181 ◽  
Author(s):  
I. Bronstein ◽  
J. Fortin ◽  
P.E. Stanley ◽  
G.S.A.B. Stewart ◽  
L.J. Kricka
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assays

Recombinant human AhR-mediated GUS reporter gene assays for PCB congeners in transgenic tobacco plants in comparison with recombinant mouse and guinea pig AhRs

2010 ◽  
Vol 45 (8) ◽  
pp. 741-749 ◽  
Author(s):  
Sayuri Shimazu ◽  
Yukiko Kawabata ◽  
Akito Inayoshi ◽  
Hideyuki Inui ◽  
Hitoshi Ashida ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Transgenic Tobacco ◽  
Reporter Gene ◽  
Transgenic Tobacco Plants ◽  
Tobacco Plants ◽  
Gus Reporter Gene ◽  
Pcb Congeners ◽  
Gus Reporter ◽  
Recombinant Mouse ◽  
Reporter Gene Assays

scholarly journals Rapid fluorescence-based reporter-gene assays to evaluate the cytotoxicity and antitumor drug potential of platinum complexes

1999 ◽  
Vol 6 (8) ◽  
pp. 541-551 ◽  
Author(s):  
Karen E Sandman ◽  
Sudhakar S Marla ◽  
Gregor Zlokarnik ◽  
Stephen J Lippard
Keyword(s):  
Reporter Gene ◽  
Platinum Complexes ◽  
Antitumor Drug ◽  
Reporter Gene Assays

Making It All Work: Functional Genomics and Reporter Gene Assays

2018 ◽  
pp. 89-105 ◽  
Author(s):  
Genevieve Welch ◽  
Robert Damoiseaux ◽  
Loren Miraglia
Keyword(s):  
Functional Genomics ◽  
Reporter Gene ◽  
Reporter Gene Assays

Reporter Gene Assays

2011 ◽  
pp. 3263-3263
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assays

scholarly journals Detection of high PBDD/Fs levels and dioxin-like activity in toys using a combination of GC-HRMS, rat-based and human-based DR CALUX® reporter gene assays

Chemosphere ◽  
2020 ◽  
Vol 251 ◽  
pp. 126579 ◽  
Author(s):  
Clémence Budin ◽  
Jindrich Petrlik ◽  
Jitka Strakova ◽  
Stephan Hamm ◽  
Bjorn Beeler ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assays

scholarly journals Regulation of differential proton-coupled folate transporter gene expression in human tumors: transactivation by KLF15 with NRF-1 and the role of Sp1

2019 ◽  
Vol 476 (8) ◽  
pp. 1247-1266
Author(s):  
Zhanjun Hou ◽  
Carrie O'Connor ◽  
Josephine Frühauf ◽  
Steve Orr ◽  
Seongho Kim ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Transcriptional Control ◽  
Consensus Sequence ◽  
Minimal Promoter ◽  
Mobility Shift ◽  
Nuclear Extracts ◽  
Ht1080 Cells ◽  
Promoter Construct ◽  
Mobility Shift Assay ◽  
Reporter Gene Assays

Abstract Tumors can be therapeutically targeted with novel antifolates (e.g. AGF94) that are selectively transported by the human proton-coupled folate transporter (hPCFT). Studies were performed to determine the transcription regulation of hPCFT in tumors and identify possible mechanisms that contribute to the highly disparate levels of hPCFT in HepG2 versus HT1080 tumor cells. Transfection of hPCFT-null HT1080 cells with hPCFT restored transport and sensitivity to AGF94. Progressive deletions of the hPCFT promoter construct (−2005 to +96) and reporter gene assays in HepG2 and HT1080 cells confirmed differences in hPCFT transactivation and localized a minimal promoter to between positions −50 and +96. The minimal promoter included KLF15, GC-Box and NRF-1 cis-binding elements whose functional importance was confirmed by promoter deletions and mutations of core consensus sequences and reporter gene assays. In HepG2 cells, NRF-1, KLF15 and Sp1 transcripts were increased over HT1080 cells by ∼5.1-, ∼44-, and ∼2.4-fold, respectively. In Drosophila SL2 cells, transfection with KLF15 and NRF-1 synergistically activated the hPCFT promoter; Sp1 was modestly activating or inhibitory. Chromatin immunoprecipitation and electrophoretic mobility shift assay (EMSA) and supershifts confirmed differential binding of KLF15, Sp1, and NRF-1 to the hPCFT promoter in HepG2 and HT1080 cells that paralleled hPCFT levels. Treatment of HT1080 nuclear extracts (NE) with protein kinase A increased Sp1 binding to its consensus sequence by EMSA, suggesting a role for Sp1 phosphorylation in regulating hPCFT transcription. A better understanding of determinants of hPCFT transcriptional control may identify new therapeutic strategies for cancer by modulating hPCFT levels in combination with hPCFT-targeted antifolates.

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