Strongly enhanced bacterial bioluminescence with the
            <i>ilux</i>
            operon for single-cell imaging

Gregor, Carola; Gwosch, Klaus; Sahl, Steffen J.; Hell, Stefan W.

doi:10.1073/pnas.1715946115

Cited by 98 publications

(110 citation statements)

References 25 publications

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“…Nonetheless, in this initial feasibility study, we estimate the glowing plants to be more than an order of magnitude brighter than previously reported using bacterial bioluminescence 24 . We anticipate that light emission could be further increased through optimizing catalytic efficacy of the fungal enzymes in plants 25,26 , improving metabolic channelling between the enzymes 27 , and enhancing precursors availability through metabolic engineering 28 .…”

Section: Discussionmentioning

confidence: 99%

Plants with self-sustained luminescence

Mitiouchkina

Mishin

Somermeyer

et al. 2019

Preprint

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In contrast to fluorescent proteins, light emission from luciferase reporters requires exogenous addition of a luciferin substrate. Bacterial bioluminescence has been the single exception, where an operon of five genes is sufficient to produce light autonomously. Although commonly used in prokaryotic hosts, toxicity of the aldehyde substrate has limited its use in eukaryotes 1 . Here we demonstrate autonomous luminescence in a multicellular eukaryotic organism by incorporating a recently discovered fungal bioluminescent system 2 into tobacco plants. We monitored these light-emitting plants from germination to flowering, observing temporal and spatial patterns of luminescence across time scales from seconds to months. The dynamic patterns of luminescence reflected progression through developmental stages, circadian oscillations, transport, and response to injuries. As with other fluorescent and luminescent reporters, we anticipate that this system will be further engineered for varied purposes, especially where exogenous addition of substrate is undesirable.

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Section: Discussionmentioning

confidence: 99%

Plants with self-sustained luminescence

Mitiouchkina

Mishin

Somermeyer

et al. 2019

Preprint

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“…Cells would thus be autoluminescent without the need for exogenous substrate (similar to the lux operon for bacterial imaging). [12] Weng and coworkers have taken key steps toward this goal by harvesting the light-emitting organs from fireflies and other insects. Transcriptome profiling revealed multiple conserved genes that are likely involved in the de novo synthesis of D-luciferin.…”

Section: Discovering New Luciferases and Luciferinsmentioning

confidence: 99%

Advances in bioluminescence imaging: new probes from old recipes

Yao

Zhang

Prescher

2018

Current Opinion in Chemical Biology

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Bioluminescent probes are powerful tools for visualizing biology in live tissues and whole animals. Recent years have seen a surge in the number of new luciferases, luciferins, and related tools available for bioluminescence imaging. Many were crafted using classic methods of optical probe design and engineering. Here we highlight recent advances in bioluminescent tool discovery and development, along with applications of the probes in cells, tissues, and organisms. Collectively, these tools are improving in vivo imaging capabilities and bolstering new research directions.

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“…As we previously reported for E. coli cells (30), the bioluminescence signal from a few HeLa cells fluctuated before cell death (Movies S3 and 4). This "blinking" occurred less frequently than in kanamycin-treated E. coli cells where it was shown to be accompanied by changes in the cellular ATP concentration (30). The cause of the blinking is unknown, but might involve similar processes affecting the energy metabolism during cell death in both cell types.…”

mentioning

confidence: 91%

Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system

Gregor

Pape

Gwosch

et al. 2019

Preprint

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Bioluminescence based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has so far not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells. Significance statementBioluminescence is generated by luciferases that oxidize a specific luciferin. The enzymes involved in the synthesis of the luciferin from widespread cellular metabolites have so far been identified for only two bioluminescence systems, those of bacteria and fungi. In these cases, the complete reaction cascade is genetically encodable, meaning that heterologous expression of the corresponding genes can potentially produce autonomous bioluminescence in cell types other than the bacterial or fungal host cells. However, the light levels achieved in mammalian cells so far are not sufficient for single-cell applications. Here we present, for the first time, autonomous bioluminescence images of single mammalian cells by coexpression of the genes encoding the six enzymes from the bacterial bioluminescence system.

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Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging

Cited by 98 publications

References 25 publications

Plants with self-sustained luminescence

Plants with self-sustained luminescence

Advances in bioluminescence imaging: new probes from old recipes

Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system

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