Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis
The bacterial bioluminescence system has been successfully implemented in mammalian cell lines, enabling the substrate-free luminescence imaging of living cells. One of the major limitations of the system is its comparatively low brightness. To improve light emission, we aimed to increase the cellul...
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MDPI AG
2024-10-01
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| Online Access: | https://www.mdpi.com/2227-9040/12/11/223 |
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| author | Theresa Brinker Carola Gregor |
| author_facet | Theresa Brinker Carola Gregor |
| author_sort | Theresa Brinker |
| collection | DOAJ |
| description | The bacterial bioluminescence system has been successfully implemented in mammalian cell lines, enabling the substrate-free luminescence imaging of living cells. One of the major limitations of the system is its comparatively low brightness. To improve light emission, we aimed to increase the cellular production of FMNH<sub>2</sub> and NADPH, which serve as cosubstrates in the bacterial bioluminescence reaction. We coexpressed different proteins involved in the synthesis of these two cofactors together with the proteins of the bacterial bioluminescence system in different mammalian cell lines. The combined expression of a riboflavin kinase (RFK) and a constitutively active Akt2 variant (Akt2CA) that participate in the cellular production of FMN and NADP<sup>+</sup>, respectively, increased bioluminescence emission up to 2.4-fold. The improved brightness allows autonomous bioluminescence imaging of mammalian cells at a higher signal-to-noise ratio and enhanced spatiotemporal resolution. |
| format | Article |
| id | doaj-art-9766e09affaa474a91f4ff9b7e026a9c |
| institution | OA Journals |
| issn | 2227-9040 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
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| series | Chemosensors |
| spelling | doaj-art-9766e09affaa474a91f4ff9b7e026a9c2025-08-20T02:08:09ZengMDPI AGChemosensors2227-90402024-10-01121122310.3390/chemosensors12110223Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor SynthesisTheresa Brinker0Carola Gregor1Department of Optical Nanoscopy, Institut für Nanophotonik Göttingen e.V., 37077 Göttingen, GermanyDepartment of Optical Nanoscopy, Institut für Nanophotonik Göttingen e.V., 37077 Göttingen, GermanyThe bacterial bioluminescence system has been successfully implemented in mammalian cell lines, enabling the substrate-free luminescence imaging of living cells. One of the major limitations of the system is its comparatively low brightness. To improve light emission, we aimed to increase the cellular production of FMNH<sub>2</sub> and NADPH, which serve as cosubstrates in the bacterial bioluminescence reaction. We coexpressed different proteins involved in the synthesis of these two cofactors together with the proteins of the bacterial bioluminescence system in different mammalian cell lines. The combined expression of a riboflavin kinase (RFK) and a constitutively active Akt2 variant (Akt2CA) that participate in the cellular production of FMN and NADP<sup>+</sup>, respectively, increased bioluminescence emission up to 2.4-fold. The improved brightness allows autonomous bioluminescence imaging of mammalian cells at a higher signal-to-noise ratio and enhanced spatiotemporal resolution.https://www.mdpi.com/2227-9040/12/11/223bioluminescencebacterial luciferasemicroscopy |
| spellingShingle | Theresa Brinker Carola Gregor Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis Chemosensors bioluminescence bacterial luciferase microscopy |
| title | Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis |
| title_full | Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis |
| title_fullStr | Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis |
| title_full_unstemmed | Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis |
| title_short | Increased Autonomous Bioluminescence Emission from Mammalian Cells by Enhanced Cofactor Synthesis |
| title_sort | increased autonomous bioluminescence emission from mammalian cells by enhanced cofactor synthesis |
| topic | bioluminescence bacterial luciferase microscopy |
| url | https://www.mdpi.com/2227-9040/12/11/223 |
| work_keys_str_mv | AT theresabrinker increasedautonomousbioluminescenceemissionfrommammaliancellsbyenhancedcofactorsynthesis AT carolagregor increasedautonomousbioluminescenceemissionfrommammaliancellsbyenhancedcofactorsynthesis |