Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development
The aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that increases the expression of detoxifying enzymes upon ligand stimulation. Recent studies now suggest that novel endogenous roles of the AHR exist throughout development. In an effort to create an optimized model syste...
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Series: | Stem Cells International |
Online Access: | http://dx.doi.org/10.1155/2016/2574152 |
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doaj-377cd6377cae445d82f756f7ca529a322020-11-25T01:08:01ZengHindawi LimitedStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/25741522574152Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human DevelopmentBrenden W. Smith0Elizabeth A. Stanford1David H. Sherr2George J. Murphy3Section of Hematology and Oncology, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USASection of Hematology and Oncology, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USADepartment of Environmental Health, Boston University School of Public Health, Boston, MA 02118, USASection of Hematology and Oncology, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USAThe aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that increases the expression of detoxifying enzymes upon ligand stimulation. Recent studies now suggest that novel endogenous roles of the AHR exist throughout development. In an effort to create an optimized model system for the study of AHR signaling in several cellular lineages, we have employed a CRISPR/CAS9 genome editing strategy in induced pluripotent stem cells (iPSCs) to incorporate a reporter cassette at the transcription start site of one of its canonical targets, cytochrome P450 1A1 (CYP1A1). This cell line faithfully reports on CYP1A1 expression, with luciferase levels as its functional readout, when treated with an endogenous AHR ligand (FICZ) at escalating doses. iPSC-derived fibroblast-like cells respond to acute exposure to environmental and endogenous AHR ligands, and iPSC-derived hepatocytes increase CYP1A1 in a similar manner to primary hepatocytes. This cell line is an important innovation that can be used to map AHR activity in discrete cellular subsets throughout developmental ontogeny. As further endogenous ligands are proposed, this line can be used to screen for safety and efficacy and can report on the ability of small molecules to regulate critical cellular processes by modulating the activity of the AHR.http://dx.doi.org/10.1155/2016/2574152 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Brenden W. Smith Elizabeth A. Stanford David H. Sherr George J. Murphy |
spellingShingle |
Brenden W. Smith Elizabeth A. Stanford David H. Sherr George J. Murphy Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development Stem Cells International |
author_facet |
Brenden W. Smith Elizabeth A. Stanford David H. Sherr George J. Murphy |
author_sort |
Brenden W. Smith |
title |
Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development |
title_short |
Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development |
title_full |
Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development |
title_fullStr |
Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development |
title_full_unstemmed |
Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development |
title_sort |
genome editing of the cyp1a1 locus in ipscs as a platform to map ahr expression throughout human development |
publisher |
Hindawi Limited |
series |
Stem Cells International |
issn |
1687-966X 1687-9678 |
publishDate |
2016-01-01 |
description |
The aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that increases the expression of detoxifying enzymes upon ligand stimulation. Recent studies now suggest that novel endogenous roles of the AHR exist throughout development. In an effort to create an optimized model system for the study of AHR signaling in several cellular lineages, we have employed a CRISPR/CAS9 genome editing strategy in induced pluripotent stem cells (iPSCs) to incorporate a reporter cassette at the transcription start site of one of its canonical targets, cytochrome P450 1A1 (CYP1A1). This cell line faithfully reports on CYP1A1 expression, with luciferase levels as its functional readout, when treated with an endogenous AHR ligand (FICZ) at escalating doses. iPSC-derived fibroblast-like cells respond to acute exposure to environmental and endogenous AHR ligands, and iPSC-derived hepatocytes increase CYP1A1 in a similar manner to primary hepatocytes. This cell line is an important innovation that can be used to map AHR activity in discrete cellular subsets throughout developmental ontogeny. As further endogenous ligands are proposed, this line can be used to screen for safety and efficacy and can report on the ability of small molecules to regulate critical cellular processes by modulating the activity of the AHR. |
url |
http://dx.doi.org/10.1155/2016/2574152 |
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