Asset Details
Do you wish to reserve the book?
Development and Application of a CAFLUX HepG2 Reporter Cell Line for Real-Time Monitoring of AhR-Mediated CYP1A1 Gene Expression in Response to Environmental Toxicants and Bioactive Modulators
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Development and Application of a CAFLUX HepG2 Reporter Cell Line for Real-Time Monitoring of AhR-Mediated CYP1A1 Gene Expression in Response to Environmental Toxicants and Bioactive Modulators
Do you wish to request the book?
Development and Application of a CAFLUX HepG2 Reporter Cell Line for Real-Time Monitoring of AhR-Mediated CYP1A1 Gene Expression in Response to Environmental Toxicants and Bioactive Modulators
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Development and Application of a CAFLUX HepG2 Reporter Cell Line for Real-Time Monitoring of AhR-Mediated CYP1A1 Gene Expression in Response to Environmental Toxicants and Bioactive Modulators
2025
Overview
This study reports the construction and validation of a CAFLUX (Chemically Activated Fluorescent Expression) HepG2 reporter cell line engineered to express a histone H2B–green fluorescent protein (H2B–GFP) fusion protein under the control of a dioxin-responsive cytochrome P450 1A1 (CYP1A1) promoter. A lentiviral construct containing a synthetic promoter with multiple dioxin-responsive elements (DREs) upstream of the H2B–EGFP coding sequence was cloned into the pFUGW vector, packaged in human embryonic kidney (HEK) 293FT cells, and used to transduce HepG2 hepatocellular carcinoma cells. Stable clones obtained by limiting dilution were screened for GFP expression in response to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The resulting CAFLUX HepG2 cells exhibited dose-dependent nuclear GFP fluorescence when exposed to aryl hydrocarbon receptor (AhR) agonists, with limits of detection of approximately 0.01 pM for TCDD and 0.1 pM for benzo[a]pyrene (B[a]P), a polycyclic aromatic hydrocarbon (PAH). This reporter activity correlated with endogenous CYP1A1 mRNA expression as determined by quantitative polymerase chain reaction (qPCR), confirming that GFP signals reflected native transcriptional responses. In functional assays, curcumin suppressed GFP expression in a concentration-dependent manner and induced apoptotic morphology at higher doses, while extracellular vesicles (EVs) derived from adipose-derived stem cells (ADSCs) significantly reduced both GFP fluorescence and CYP1A1 mRNA levels, suggesting an inhibitory effect on AhR-driven transcription. The CAFLUX HepG2 reporter system therefore provides a sensitive and reproducible platform for real-time, nuclear-localized monitoring of AhR-mediated gene expression. Its responsiveness to both agonists and antagonists underscores its potential utility in toxicological evaluation, drug discovery, and the investigation of EV-mediated signaling in liver cancer models.
Publisher
MDPI AG
Subject
Basic Helix-Loop-Helix Transcription Factors
/ Cytochrome P-450 CYP1A1 - genetics
/ Cytochrome P-450 CYP1A1 - metabolism
/ Dioxin
/ Dioxins
/ Environmental Pollutants - toxicity
/ Enzymes
/ Genes
/ Green Fluorescent Proteins - genetics
/ Green Fluorescent Proteins - metabolism
/ Humans
/ Ligands
/ Plasmids
/ Polychlorinated Dibenzodioxins - pharmacology
/ Polychlorinated Dibenzodioxins - toxicity
/ Receptors, Aryl Hydrocarbon - genetics
/ Receptors, Aryl Hydrocarbon - metabolism
/ RNA
/ Toxicity
