Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.

Usage of the enhanced green fluorescent protein (eGFP) in living mammalian cells is limited to aerobic conditions due to requirement of oxygen during chromophore formation. Since many diseases or disease models are associated with acute or chronic hypoxia, eGFP-labeling of structures of interest in...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Janine Walter, Sascha Hausmann, Thomas Drepper, Michael Puls, Thorsten Eggert, Marcel Dihné
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/db363103e0584e55b89d298090ce9e00
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:db363103e0584e55b89d298090ce9e00
record_format	dspace
spelling	oai:doaj.org-article:db363103e0584e55b89d298090ce9e002021-11-18T07:06:06ZFlavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.1932-620310.1371/journal.pone.0043921https://doaj.org/article/db363103e0584e55b89d298090ce9e002012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22984451/?tool=EBIhttps://doaj.org/toc/1932-6203Usage of the enhanced green fluorescent protein (eGFP) in living mammalian cells is limited to aerobic conditions due to requirement of oxygen during chromophore formation. Since many diseases or disease models are associated with acute or chronic hypoxia, eGFP-labeling of structures of interest in experimental studies might be unreliable leading to biased results. Thus, a chromophore yielding a stable fluorescence under hypoxic conditions is desirable. The fluorescence of flavin mononucleotide (FMN)-based fluorescent proteins (FbFPs) does not require molecular oxygen. Recently, the advantages of FbFPs for several bacterial strains and yeasts were described, specifically, their usage as a real time fluorescence marker in bacterial expression studies and their ability of chromophore formation under anaerobic conditions. Our objective was to verify if FbFPs also function in mammalian cells in order to potentially broaden the repertoire of chromophores with ones that can reliably be used in mammalian studies under hypoxic conditions. In the present study, we demonstrate for the first time, that FbFPs can be expressed in different mammalian cells, among them murine neural stem cells during proliferative and differentiated stages. Fluorescence intensities were comparable to eGFP. In contrast to eGFP, the FbFP fluorescence did not decrease when cells were exposed to defined hypoxic conditions neither in proliferating nor in differentiated cells. Thus, FbFPs can be regarded as an alternative to eGFP in studies that target cellular structures which are exposed to hypoxic conditions.Janine WalterSascha HausmannThomas DrepperMichael PulsThorsten EggertMarcel DihnéPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 9, p e43921 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Janine Walter Sascha Hausmann Thomas Drepper Michael Puls Thorsten Eggert Marcel Dihné Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
description	Usage of the enhanced green fluorescent protein (eGFP) in living mammalian cells is limited to aerobic conditions due to requirement of oxygen during chromophore formation. Since many diseases or disease models are associated with acute or chronic hypoxia, eGFP-labeling of structures of interest in experimental studies might be unreliable leading to biased results. Thus, a chromophore yielding a stable fluorescence under hypoxic conditions is desirable. The fluorescence of flavin mononucleotide (FMN)-based fluorescent proteins (FbFPs) does not require molecular oxygen. Recently, the advantages of FbFPs for several bacterial strains and yeasts were described, specifically, their usage as a real time fluorescence marker in bacterial expression studies and their ability of chromophore formation under anaerobic conditions. Our objective was to verify if FbFPs also function in mammalian cells in order to potentially broaden the repertoire of chromophores with ones that can reliably be used in mammalian studies under hypoxic conditions. In the present study, we demonstrate for the first time, that FbFPs can be expressed in different mammalian cells, among them murine neural stem cells during proliferative and differentiated stages. Fluorescence intensities were comparable to eGFP. In contrast to eGFP, the FbFP fluorescence did not decrease when cells were exposed to defined hypoxic conditions neither in proliferating nor in differentiated cells. Thus, FbFPs can be regarded as an alternative to eGFP in studies that target cellular structures which are exposed to hypoxic conditions.
format	article
author	Janine Walter Sascha Hausmann Thomas Drepper Michael Puls Thorsten Eggert Marcel Dihné
author_facet	Janine Walter Sascha Hausmann Thomas Drepper Michael Puls Thorsten Eggert Marcel Dihné
author_sort	Janine Walter
title	Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
title_short	Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
title_full	Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
title_fullStr	Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
title_full_unstemmed	Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
title_sort	flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/db363103e0584e55b89d298090ce9e00
work_keys_str_mv	AT janinewalter flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement AT saschahausmann flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement AT thomasdrepper flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement AT michaelpuls flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement AT thorsteneggert flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement AT marceldihne flavinmononucleotidebasedfluorescentproteinsfunctioninmammaliancellswithoutoxygenrequirement
_version_	1718423947893014528

Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.

Ejemplares similares