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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22984451/?tool=EBI |
| _version_ | 1818434959272050688 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-14T16:45:16Z |
| format | Article |
| id | doaj.art-db363103e0584e55b89d298090ce9e00 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T16:45:16Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-db363103e0584e55b89d298090ce9e002022-12-21T22:54:12ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0179e4392110.1371/journal.pone.0043921Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement.Janine WalterSascha HausmannThomas DrepperMichael PulsThorsten EggertMarcel DihnéUsage 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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22984451/?tool=EBI |
| spellingShingle | Janine Walter Sascha Hausmann Thomas Drepper Michael Puls Thorsten Eggert Marcel Dihné Flavin mononucleotide-based fluorescent proteins function in mammalian cells without oxygen requirement. PLoS ONE |
| title | 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_short | 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 |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22984451/?tool=EBI |
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