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Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells

A unique hybrid protein ferritin–enhanced green fluorescent protein (EGFP) was built to serve as an endogenous dual reporter for both fluorescence and magnetic resonance imaging (MRI). It consists of a human ferritin heavy chain (an iron-storage protein) at the N terminus, a flexible polypeptide in...

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Detalles Bibliográficos
Autores principales: Jiang, Caihong, Wu, Dongmei, Haacke, E. Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Grapho Publications, LLC 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6024424/
https://www.ncbi.nlm.nih.gov/pubmed/30042970
http://dx.doi.org/10.18383/j.tom.2015.00181
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author Jiang, Caihong
Wu, Dongmei
Haacke, E. Mark
author_facet Jiang, Caihong
Wu, Dongmei
Haacke, E. Mark
author_sort Jiang, Caihong
collection PubMed
description A unique hybrid protein ferritin–enhanced green fluorescent protein (EGFP) was built to serve as an endogenous dual reporter for both fluorescence and magnetic resonance imaging (MRI). It consists of a human ferritin heavy chain (an iron-storage protein) at the N terminus, a flexible polypeptide in the middle as a linker, and an EGFP at the C terminus. Through antibiotic screening, we established stable human glioma U251 cell strains that expressed ferritin–EGFP under the control of tetracycline. These cells emitted bright green fluorescence and were easily detected by a fluorescent microscope. Ferritin–EGFP overexpression proved effective in triggering obvious intracellular iron accumulation as shown by Prussian blue staining and by MRI. Further, we found that ferritin–EGFP overexpression did not cause proliferation differences between experimental and control group cells when ferritin–EGFP was expressed for <96 hours. Application of this novel ferritin–EGFP chimera has a promising future for combined optical and MRI approaches to study in vivo imaging at a cellular level.
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spelling pubmed-60244242018-07-24 Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells Jiang, Caihong Wu, Dongmei Haacke, E. Mark Tomography Advances in Brief A unique hybrid protein ferritin–enhanced green fluorescent protein (EGFP) was built to serve as an endogenous dual reporter for both fluorescence and magnetic resonance imaging (MRI). It consists of a human ferritin heavy chain (an iron-storage protein) at the N terminus, a flexible polypeptide in the middle as a linker, and an EGFP at the C terminus. Through antibiotic screening, we established stable human glioma U251 cell strains that expressed ferritin–EGFP under the control of tetracycline. These cells emitted bright green fluorescence and were easily detected by a fluorescent microscope. Ferritin–EGFP overexpression proved effective in triggering obvious intracellular iron accumulation as shown by Prussian blue staining and by MRI. Further, we found that ferritin–EGFP overexpression did not cause proliferation differences between experimental and control group cells when ferritin–EGFP was expressed for <96 hours. Application of this novel ferritin–EGFP chimera has a promising future for combined optical and MRI approaches to study in vivo imaging at a cellular level. Grapho Publications, LLC 2017-03 /pmc/articles/PMC6024424/ /pubmed/30042970 http://dx.doi.org/10.18383/j.tom.2015.00181 Text en © 2017 The Authors. Published by Grapho Publications, LLC http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Advances in Brief
Jiang, Caihong
Wu, Dongmei
Haacke, E. Mark
Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title_full Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title_fullStr Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title_full_unstemmed Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title_short Ferritin–EGFP Chimera as an Endogenous Dual-Reporter for Both Fluorescence and Magnetic Resonance Imaging in Human Glioma U251 Cells
title_sort ferritin–egfp chimera as an endogenous dual-reporter for both fluorescence and magnetic resonance imaging in human glioma u251 cells
topic Advances in Brief
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6024424/
https://www.ncbi.nlm.nih.gov/pubmed/30042970
http://dx.doi.org/10.18383/j.tom.2015.00181
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