Red-shifted click beetle luciferase mutant expands the multicolor bioluminescent palette for deep tissue imaging

Zambito, Giorgia; Hall, Mary P.; Wood, Monika G.; Gaspar, Natasa; Ridwan, Yanto; Stellari, Fabio; Shi, Ce; Kirkland, Thomas A.; Encell, Lance P.; Löwik, Clemens; Mezzanotte, Laura

doi:10.1016/j.isci.2020.101986

Cited by 34 publications

(39 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fluorescent proteins are used as intracellular gene reporters, biomarkers of protein expression or of dynamic signaling events and trackers of targeted cells, as in tumor invasion or cell transplantation even in deep tissues as brain or lung, in in vivo experimental animal [257][258][259][260][261][262]. The main fluorophores described in this section are summarized in Table 11.…”

Section: Bioluminescencementioning

confidence: 99%

Light and Autofluorescence, Multitasking Features in Living Organisms

Croce

2021

Photochem

View full text Add to dashboard Cite

Organisms belonging to all life kingdoms may have the natural capacity to fluoresce. Autofluorescence events depend on the presence of natural biomolecules, namely endogenous fluorophores, with suitable chemical properties in terms of conjugated double bonds, aromatic or more complex structures with oxidized and crosslinked bonds, ensuring an energy status able to permit electronic transitions matching with the energy of light in the UV-visible-near-IR spectral range. Emission of light from biological substrates has been reported since a long time, inspiring unceasing and countless studies. Early notes on autofluorescence of vegetables have been soon followed by attention to animals. Investigations on full living organisms from the wild environment have been driven prevalently by ecological and taxonomical purposes, while studies on cells, tissues and organs have been mainly promoted by diagnostic aims. Interest in autofluorescence is also growing as a sensing biomarker in food production and in more various industrial processes. The associated technological advances have supported investigations ranging from the pure photochemical characterization of specific endogenous fluorophores to their possible functional meanings and biological relevance, making fluorescence a valuable intrinsic biomarker for industrial and diagnostic applications, in a sort of real time, in situ biochemical analysis. This review aims to provide a wide-ranging report on the most investigated natural fluorescing biomolecules, from microorganisms to plants and animals of different taxonomic degrees, with their biological, environmental or biomedical issues relevant for the human health. Hence, some notes in the different sections dealing with different biological subject are also interlaced with human related issues. Light based events in biological subjects have inspired an almost countless literature, making it almost impossible to recall here all associated published works, forcing to apologize for the overlooked reports. This Review is thus proposed as an inspiring source for Readers, addressing them to additional literature for an expanded information on specific topics of more interest.

show abstract

Section: Bioluminescencementioning

confidence: 99%

Light and Autofluorescence, Multitasking Features in Living Organisms

Croce

2021

Photochem

View full text Add to dashboard Cite

show abstract

“…9,10 Multiplexed imaging is possible using spectrally resolved luciferase-luciferin pairs. [11][12][13][14] Indeed, several luciferin analogs have been developed that emit different colors of light. [15][16][17] The emission proles can be further tuned with engineered luciferases 18,19 or luciferase-uorescent probe fusions.…”

Section: Introductionmentioning

confidence: 99%

Coumarin luciferins and mutant luciferases for robust multi-component bioluminescence imaging

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Luc146-1H2 is a thermostable variant created by mutating the naturally occurring beetle luciferase ( LucPpe). Multiple studies have shown that specific mutations in luciferases will shift their emission spectrum, therefore, further studies are needed to check whether the Luc146-1H2 mutant would led to a change in emission spectrum in comparison to LucPpe that can be potentially exploited for multicolor in vivo BLI imaging 24 – 27 .…”

Section: Discussionmentioning

confidence: 99%

A novel thermostable beetle luciferase based cytotoxicity assay

Choi

Matta

Gopalakrishnan

et al. 2021

Sci Rep

View full text Add to dashboard Cite

Cytotoxicity assays are essential for the testing and development of novel immunotherapies for the treatment of cancer. We recently described a novel cytotoxicity assay, termed the Matador assay, which was based on marine luciferases and their engineered derivatives. In this study, we describe the development of a new cytotoxicity assay termed ‘Matador-Glo assay’ which takes advantage of a thermostable variant of Click Beetle Luciferase (Luc146-1H2). Matador-Glo assay utilizes Luc146-1H2 and D-luciferin as the luciferase-substrate pair for luminescence detection. The assay involves ectopic over-expression of Luc146-1H2 in the cytosol of target cells of interest. Upon damage to the membrane integrity, the Luc146-1H2 is either released from the dead and dying cells or its activity is preferentially measured in dead and dying cells. We demonstrate that this assay is simple, fast, specific, sensitive, cost-efficient, and not labor-intensive. We further demonstrate that the Matador-Glo assay can be combined with the marine luciferase-based Matador assay to develop a dual luciferase assay for cell death detection. Finally, we demonstrate that the Luc146-1H2 expressing target cells can also be used for in vivo bioluminescence imaging applications.

show abstract

Red-shifted click beetle luciferase mutant expands the multicolor bioluminescent palette for deep tissue imaging

Cited by 34 publications

References 26 publications

Light and Autofluorescence, Multitasking Features in Living Organisms

Light and Autofluorescence, Multitasking Features in Living Organisms

Coumarin luciferins and mutant luciferases for robust multi-component bioluminescence imaging

A novel thermostable beetle luciferase based cytotoxicity assay

Contact Info

Product

Resources

About