Near infrared bioluminescence resonance energy transfer from firefly luciferase—quantum dot bionanoconjugates

Alam, Rabeka; Karam, Liliana M; Doane, Tennyson L.; Zylstra, Joshua; Fontaine, Danielle M.; Branchini, Bruce R.; Maye, Mathew M.

doi:10.1088/0957-4484/25/49/495606

Cited by 30 publications

(26 citation statements)

References 34 publications

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“…20,21 In practice, bioluminescing proteins such as Renilla reniformis luciferase (Luc) are attached to the QDs where their oxidation of a chemical substrate produces either localized light emission or an exciton that can sensitize the QD. 29,30 More pertinently, they showed that attaching red-shifted fluorescent proteins to the QDs could give rise to a multistep BRET-FRET process; this served as an important confirmation from which we would build upon. [21][22][23][24] Quinones et al extended this QD assay format to quantitatively probe cellular surface-and proteinprotein interactions in a manner that was far more sensitive than conventional direct fluorescence-based approaches.…”

Section: Introductionmentioning

confidence: 70%

An enzymatically-sensitized sequential and concentric energy transfer relay self-assembled around semiconductor quantum dots

et al. 2015

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The ability to control light energy within de novo nanoscale structures and devices will greatly benefit their continuing development and ultimate application. Ideally, this control should extend from generating the light itself to its spatial propagation within the device along with providing defined emission wavelength(s), all in a stand-alone modality. Here we design and characterize macromolecular nanoassemblies consisting of semiconductor quantum dots (QDs), several differentially dye-labeled peptides and the enzyme luciferase which cumulatively demonstrate many of these capabilities by engaging in multiple-sequential energy transfer steps. To create these structures, recombinantly-expressed luciferase and the dye-labeled peptides were appended with a terminal polyhistidine sequence allowing for controlled ratiometric self-assembly around the QDs via metal-affinity coordination. The QDs serve to provide multiple roles in these structures including as central assembly platforms or nanoscaffolds along with acting as a potent energy harvesting and transfer relay. The devices are activated by addition of coelenterazine H substrate which is oxidized by luciferase producing light energy which sensitizes the central 625 nm emitting QD acceptor by bioluminescence resonance energy transfer (BRET). The sensitized QD, in turn, acts as a relay and transfers the energy to a first peptide-labeled Alexa Fluor 647 acceptor dye displayed on its surface. This dye then transfers energy to a second red-shifted peptide-labeled dye acceptor on the QD surface through a second concentric Förster resonance energy transfer (FRET) process. Alexa Fluor 700 and Cy5.5 are both tested in the role of this terminal FRET acceptor. Photophysical analysis of spectral profiles from the resulting sequential BRET-FRET-FRET processes allow us to estimate the efficiency of each of the transfer steps. Importantly, the efficiency of each step within this energy transfer cascade can be controlled to some extent by the number of enzymes/peptides displayed on the QD. Further optimization of the energy transfer process(es) along with potential applications of such devices are finally discussed.

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Section: Introductionmentioning

confidence: 70%

An enzymatically-sensitized sequential and concentric energy transfer relay self-assembled around semiconductor quantum dots

et al. 2015

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“…The incorporation of luciferase-encoding plasmids into bacteria is a common approach for the development of bioluminescent reporter strains (26). Chromosomally integrated luciferases are often preferred (27), because they are more stable than plasmid-based reporters, although sensitivity is generally lower (17,28). It is recognized that plasmid-based reporters may be unstable in vivo in the absence of antibiotic selection (23,29,30).…”

Section: Discussionmentioning

confidence: 99%

A Bioluminescent Francisella tularensis SCHU S4 Strain Enables Noninvasive Tracking of Bacterial Dissemination and the Evaluation of Antibiotics in an Inhalational Mouse Model of Tularemia

Hall

Flick-Smith

Harding

et al. 2016

Antimicrob Agents Chemother

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bBioluminescence imaging (BLI) enables real-time, noninvasive tracking of infection in vivo and longitudinal infection studies. In this study, a bioluminescent Francisella tularensis strain, SCHU S4-lux, was used to develop an inhalational infection model in BALB/c mice. Mice were infected intranasally, and the progression of infection was monitored in real time using BLI. A bioluminescent signal was detectable from 3 days postinfection (3 dpi), initially in the spleen and then in the liver and lymph nodes, before finally becoming systemic. The level of bioluminescent signal correlated with bacterial numbers in vivo, enabling noninvasive quantification of bacterial burdens in tissues. Treatment with levofloxacin (commencing at 4 dpi) significantly reduced the BLI signal. Furthermore, BLI was able to distinguish noninvasively between different levofloxacin treatment regimens and to identify sites of relapse following treatment cessation. These data demonstrate that BLI and SCHU S4-lux are suitable for the study of F. tularensis pathogenesis and the evaluation of therapeutics for tularemia. Francisella tularensis is a Gram-negative intracellular bacterium and is the etiological agent of the disease tularemia. Tularemia has a low infectious dose by the aerosol route (50% infectious dose [ID 50 ], Ͻ10 CFU), causing a disabling illness without rapid treatment, and is considered a potential biothreat agent. The preferred treatments for tularemia are limited to a few antibiotics, including fluoroquinolones (1, 2), tetracycline (3), streptomycin (4, 5), and gentamicin (6, 7). The lack of a licensed vaccine, reported treatment failures, even with preferred antibiotics (8), and a 2% mortality rate despite treatment (5) necessitate the development and evaluation of new therapies.Conventional methods of assessing bacterial growth and dissemination in vivo require the infection of large cohorts of animals and the culling of groups of animals at set time points throughout a study. This strategy not only uses large numbers of animals but is also labor-intensive and subject to tissue sampling bias. Noninvasive imaging techniques, such as bioluminescence imaging (BLI), can report spatial and temporal aspects of infectious diseases in vivo both longitudinally and in real time (9). BLI typically relies on the detection of light produced by luciferase-catalyzed oxidation reactions, which are encoded in the pathogen of interest, by use of a sensitive charge-coupled device (CCD) camera (10). Pivotal experiments by Contag et al. have demonstrated the usefulness of BLI in the noninvasive study of bacterial pathogenesis and the evaluation of antibiotic treatments for Salmonella enterica serovar Typhimurium (11). Subsequently, bioluminescent reporters have been introduced into other bacteria, including Mycobacterium tuberculosis (10, 12), Staphylococcus aureus (13, 14), Burkholderia mallei (15), and Yersinia pestis (16,17). Further, BLI has been used to evaluate vaccines (18), antibiotics (19-21), and supportive therapies (22) for a ran...

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“…In parallel to the development of optimized BRET efficiency, the sophistication of such systems has steadily grown by integrating other elements, such as organic fluorophores [57] or FPs [58] to the existing constructs. They can work in concert with the QD to harvest luciferase light energy and utilize it by spatially and spectrally propagating it in a unique fashion that can potentially broaden their breadth of application.…”

Section: Bret-multistep Fret Constructsmentioning

confidence: 99%

Bioluminescence-Based Energy Transfer Using Semiconductor Quantum Dots as Acceptors

Samanta

Medintz

2020

Sensors

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Bioluminescence resonance energy transfer (BRET) is the non-radiative transfer of energy from a bioluminescent protein donor to a fluorophore acceptor. It shares all the formalism of Förster resonance energy transfer (FRET) but differs in one key aspect: that the excited donor here is produced by biochemical means and not by an external illumination. Often the choice of BRET source is the bioluminescent protein Renilla luciferase, which catalyzes the oxidation of a substrate, typically coelenterazine, producing an oxidized product in its electronic excited state that, in turn, couples with a proximal fluorophore resulting in a fluorescence emission from the acceptor. The acceptors pertinent to this discussion are semiconductor quantum dots (QDs), which offer some unrivalled photophysical properties. Amongst other advantages, the QD’s large Stokes shift is particularly advantageous as it allows easy and accurate deconstruction of acceptor signal, which is difficult to attain using organic dyes or fluorescent proteins. QD-BRET systems are gaining popularity in non-invasive bioimaging and as probes for biosensing as they don’t require external optical illumination, which dramatically improves the signal-to-noise ratio by avoiding background auto-fluorescence. Despite the additional advantages such systems offer, there are challenges lying ahead that need to be addressed before they are utilized for translational types of research.

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Near infrared bioluminescence resonance energy transfer from firefly luciferase—quantum dot bionanoconjugates

Cited by 30 publications

References 34 publications

An enzymatically-sensitized sequential and concentric energy transfer relay self-assembled around semiconductor quantum dots

An enzymatically-sensitized sequential and concentric energy transfer relay self-assembled around semiconductor quantum dots

A Bioluminescent Francisella tularensis SCHU S4 Strain Enables Noninvasive Tracking of Bacterial Dissemination and the Evaluation of Antibiotics in an Inhalational Mouse Model of Tularemia

Bioluminescence-Based Energy Transfer Using Semiconductor Quantum Dots as Acceptors

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