ATP-Independent Bioluminescent Reporter Variants To Improve <i>in Vivo</i> Imaging

Yeh, Hsien‐Wei; Xiong, Ying; Wu, Tianchen; Chen, Minghai; Ji, Ao; Li, Xinyu; Ai, Hui‐wang

doi:10.1021/acschembio.9b00150

Cited by 57 publications

(75 citation statements)

References 32 publications

(65 reference statements)

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“…Of note, other efforts to improve the sensitivity of BLI for in vivo imaging include directed evolution of luciferase from marine organisms, which typically emit blue light, such as Nanoluc, for alternative substrates that emit light of longer wave lengths, or the coupling of blue-emitting BLI systems with orange fluorescent proteins for FRET effects. 12 , 13 Such approaches may potentially reach in vivo sensitivity similar to that of Akaluc BLI and represent useful additions due to their distinct substrate specificities, thus allowing 2-population BLI. 13 …”

Section: Discussionmentioning

confidence: 99%

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Bozec

Sattiraju

Bouras

et al. 2020

Neuro-Oncology Advances

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Background Longitudinal tracking of tumor growth using non-invasive bioluminescence imaging (BLI) is a key approach for studies of in vivo cancer models, with particular relevance for investigations of malignant gliomas in rodent intracranial transplant paradigms. Akaluciferase (Akaluc) is a new BLI system with higher signal strength than standard firefly luciferase (Fluc). Here, we establish Akaluc BLI as a sensitive method for in vivo tracking of glioma expansion. Methods We engineered a lentiviral vector for expression of Akaluc in high-grade glioma cell lines, including patient-derived glioma stem cell (GSC) lines. Akaluc expressing glioma cells were compared to matching cells expressing Fluc in both in vitro and in vivo BLI assays. We also conducted proof-of-principle BLI studies with intracranial transplant cohorts receiving chemo-radiation therapy. Results Akaluc-expressing glioma cells produced more than 10 times higher BLI signals than Fluc-expressing counterparts when examined in vitro, and more than 100-fold higher signals when compared to Fluc-expressing counterparts in intracranial transplant models in vivo. The high sensitivity of Akaluc permitted detection of intracranial glioma transplants starting as early as 4 hours after implantation and with as little as 5,000 transplanted cells. The sensitivity of the system allowed us to follow engraftment and expansion of intracranial transplants of GSC lines. Akaluc was also robust for sensitive detection of in vivo tumor regression after therapy and subsequent relapse. Conclusion Akaluc BLI offers superior sensitivity for in vivo tracking of glioma in the intracranial transplant paradigm, facilitating sensitive approaches for the study of glioma growth and response to therapy.

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Section: Discussionmentioning

confidence: 99%

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Bozec

Sattiraju

Bouras

et al. 2020

Neuro-Oncology Advances

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“…Freed from the limited luciferin repertoire that exists in nature, synthetic luciferin analogues are helping to expand the bioluminescent intersection with classes of enzymes that share homology with luciferases . Studies in this direction also have considerable practical significance for bioluminescence imaging applications .…”

Section: Discussionmentioning

confidence: 99%

“…) . While in‐depth discussion of these and related luciferins is beyond the scope of this review , it is instructive to contrast luciferases that utilize coelenterazine as their substrate to luciferases that use d ‐luciferin as substrate. A number of coelenterazine‐utilizing luciferases are well known as they have been repurposed for laboratory use (e.g., luciferases from Renilla , Gaussia , and Oplophorus ) .…”

Section: Luciferases and Luciferins From Other Bioluminescent Organismsmentioning

confidence: 99%

Enzymatic promiscuity and the evolution of bioluminescence

Adams

Miller

2019

The FEBS Journal

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Bioluminescence occurs when an enzyme, known as a luciferase, oxidizes a small‐molecule substrate, known as a luciferin. Nature has evolved multiple distinct luciferases and luciferins independently, all of which accomplish the impressive feat of light emission. One of the best‐known examples of bioluminescence is exhibited by fireflies, a class of beetles that use d‐luciferin as their substrate. The evolution of bioluminescence in beetles is thought to have emerged from ancestral fatty acyl‐CoA synthetase (ACS) enzymes present in all insects. This theory is supported by multiple lines of evidence: Beetle luciferases share high sequence identity with these enzymes, often retain ACS activity, and some ACS enzymes from nonluminous insects can catalyze bioluminescence from synthetic d‐luciferin analogues. Recent sequencing of firefly genomes and transcriptomes further illuminates how the duplication of ACS enzymes and subsequent diversification drove the evolution of bioluminescence. These genetic analyses have also uncovered candidate enzymes that may participate in luciferin metabolism. With the publication of the genomes and transcriptomes of fireflies and related insects, we are now better positioned to dissect and learn from the evolution of bioluminescence in beetles.

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“…In living mouse organs, on the other hand, Antares2 shows an additional 35–90% signal increase over teLuc, which again shows that the redder one is better for deep tissue BLI. The engineered luciferase LumiLuc improved from teLuc with mutations at E4G, L18Q, S19A, V27L, S28T, G67C, G71A, N85D, V90A, R112Q, V119K, and K136T, was also exploited as a BRET donor [ 25 ]. The BRET pair between LumiLuc and mScarlet with 8pyDTZ ( Figure 3 k) used as a substrate emits bioluminescence around 600 nm.…”

Section: Development Of Red-shifted Bioluminescence Systemmentioning

confidence: 99%

Advanced Bioluminescence System for In Vivo Imaging with Brighter and Red-Shifted Light Emission

Endo

Ozawa

2020

IJMS

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In vivo bioluminescence imaging (BLI), which is based on luminescence emitted by the luciferase–luciferin reaction, has enabled continuous monitoring of various biochemical processes in living animals. Bright luminescence with a high signal-to-background ratio, ideally red or near-infrared light as the emission maximum, is necessary for in vivo animal experiments. Various attempts have been undertaken to achieve this goal, including genetic engineering of luciferase, chemical modulation of luciferin, and utilization of bioluminescence resonance energy transfer (BRET). In this review, we overview a recent advance in the development of a bioluminescence system for in vivo BLI. We also specifically examine the improvement in bioluminescence intensity by mutagenic or chemical modulation on several beetle and marine luciferase bioluminescence systems. We further describe that intramolecular BRET enhances luminescence emission, with recent attempts for the development of red-shifted bioluminescence system, showing great potency in in vivo BLI. Perspectives for future improvement of bioluminescence systems are discussed.

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ATP-Independent Bioluminescent Reporter Variants To Improve in Vivo Imaging

Cited by 57 publications

References 32 publications

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Akaluc bioluminescence offers superior sensitivity to track in vivo glioma expansion

Enzymatic promiscuity and the evolution of bioluminescence

Advanced Bioluminescence System for In Vivo Imaging with Brighter and Red-Shifted Light Emission

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