Advances in bioluminescence imaging: new probes from old recipes

Yao, Zi; Zhang, Brendan S.; Prescher, Jennifer A.

doi:10.1016/j.cbpa.2018.05.009

Cited by 94 publications

(74 citation statements)

References 58 publications

(69 reference statements)

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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%

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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Section: Discussionmentioning

confidence: 99%

Enzymatic promiscuity and the evolution of bioluminescence

Adams

Miller

2019

The FEBS Journal

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“…To reduce variability of measurements as much as possible, stable cell lines could be generated by the targeted integration of all multiplex reporters at the same defined site in the genome, such as a safe harbor site 50 , resulting in the neutralization of genomic position effects. Cells with such genome-integrated luciferase reporters can then be further used to explore luciferase multiplexing towards in vivo bioluminescence applications [51][52][53] . Another concern may be potential cross-talk between the reporters integrated into the same plasmid by synthetic biology.…”

Section: Discussionmentioning

confidence: 99%

Examining multiple cellular pathways at once using multiplex hextuple luciferase assaying

Sarrión-Perdigones

Chang

Gonzalez

et al. 2019

Preprint

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Sensitive simultaneous assessment of multiple signaling pathways within the same cells requires orthogonal reporters that can assay over large dynamic ranges. Luciferases have arisen as possible genetically encoded candidates due to their sensitivity, versatility, and cost-effectiveness. Here, we expanded luciferase multiplexing in post-lysis endpoint luciferase assays from two towards six. Light emissions are distinguished by a combination of distinct substrates and emission spectra deconvolution. Using synthetic assembly cloning, all six luciferase reporter units are stitched together into one plasmid; facilitating delivery of all reporter units through a process we named solotransfection, minimizing experimental errors. We engineered a multiplex hextuple luciferase assay to probe pathway fluxes through five transcriptional response elements against a control constitutive promoter. We were able to monitor the effects of siRNA, ligand, and chemical compound treatments on their target pathways along with the four other probed cellular pathways.We demonstrate the effectiveness and adaptiveness of multiplex luciferase assaying, as well as its broad application across different research fields.Running title: Multiplex hextuple luciferase assaying -3-

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“…Water absorbs at longer wavelengths [44]. Luminescence based on cellular expression of luciferase enzymes converting substrates to visible light gives superior images because it avoids illumination and corresponding tissue autofluorescence [45]. Fluorescent images are co-registered to hybrid images using brightfield or X-ray anatomical imaging.…”

Section: Preclinical Hybrid Imagingmentioning

confidence: 99%