A redshifted codon-optimized firefly luciferase is a sensitive reporter for bioluminescence imaging

Caysa, Henrike; Jacob, Roland; Müther, Nadine; Branchini, Bruce R.; Messerle, Martin; Söling, Ariane

doi:10.1039/b814566k

Cited by 48 publications

(38 citation statements)

References 31 publications

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“…Data reported the mean percentage of six different red and green tumors. This result is in agreement with previous evidence that light emitted from red-shifted luciferase passes through tissues three times more efficiently than does green light [13].…”

Section: Scarification Studies To Compare Red and Green Signal Absorpsupporting

confidence: 93%

“…The whole animal BL emission from the HepG2-Ppyred cells was much higher than that obtained from HepG2-Luc, with a mean recovery of the red photons 2:3 AE 0:3 Â 10 5 ph=s=mm 2 ð Þ three times higher than that of the green photons 7:8 AE 0:3 Â 10 4 ph=s=mm 2 ð Þ . These results are consistent with previous experiments, wherein mice were injected with nude plasmids encoding WT luciferase and a red-emitting luciferase (S284T) [13].…”

Section: Bli Of the Hepatoblastoma (Hepg2) Xenograft Murine Model In supporting

confidence: 93%

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In Vivo Bioluminescence Imaging of Murine Xenograft Cancer Models with a Red-shifted Thermostable Luciferase

et al. 2009

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Two different bioluminescent mouse cancer models demonstrate the utility of a new red-shifted thermostable luciferase, Ppy RE-TS, that improved the in vivo imaging performance when compared with wild-type P. Pyralis luciferase. While wild-type luciferase is currently a popular reporter for in vivo imaging methods, this study demonstrates the potential of red-emitting firefly luciferase mutants to enhance the performance of bioluminescence imaging experiments.

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Section: Scarification Studies To Compare Red and Green Signal Absorpsupporting

confidence: 93%

Section: Bli Of the Hepatoblastoma (Hepg2) Xenograft Murine Model In supporting

confidence: 93%

In Vivo Bioluminescence Imaging of Murine Xenograft Cancer Models with a Red-shifted Thermostable Luciferase

et al. 2009

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“…Reprinted with permission (43). modality (7,14). Additionally, alternative methods for transduction of reporter gene may reduce the risk of anomalous and inappropriate transcription of unwanted sequences in mammalian cells (37).…”

Section: Discussionmentioning

confidence: 99%

In vivo bioluminescence for tracking cell fate and function

Almeida¹,

Rappard

2011

American Journal of Physiology-Heart and Circulatory Physiology

101

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Tracking the fate and function of cells in vivo is paramount for the development of rational therapies for cardiac injury. Bioluminescence imaging (BLI) provides a means for monitoring physiological processes in real time, ranging from cell survival to gene expression to complex molecular processes. In mice and rats, BLI provides unmatched sensitivity because of the absence of endogenous luciferase expression in mammalian cells and the low background luminescence emanating from animals. In the field of stem cell therapy, BLI provides an unprecedented means to monitor the biology of these cells in vivo, giving researchers a greater understanding of their survival, migration, immunogenicity, and potential tumorigenicity in a living animal. In addition to longitudinal monitoring of cell survival, BLI is a useful tool for semiquantitative measurements of gene expression in vivo, allowing a better optimization of drug and gene therapies. Overall, this technology not only enables rapid, reproducible, and quantitative monitoring of physiological processes in vivo but also can measure the influences of therapeutic interventions on the outcome of cardiac injuries.

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“…We recently found that human codon optimization of a related red light-emitting luciferase led to a 10-fold improvement in luciferase detectability in the HEK293 cell line (6). The stability of the bioluminescence signal could be improved by insertion of the luciferase reporter gene in the chromosome (20).…”

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confidence: 99%

In Vivo Bioluminescence Imaging for the Study of Intestinal Colonization by Escherichia coli in Mice

Foucault

Thomas

Goussard

et al. 2010

Appl Environ Microbiol

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Bioluminescence imaging (BLI) is emerging as a powerful tool for real-time monitoring of infections in living animals. However, since luciferases are oxygenases, it has been suggested that the requirement for oxygen may limit the use of BLI in anaerobic environments, such as the lumen of the gut. Strains of Escherichia coli harboring the genes for either the bacterial luciferase from Photorhabdus luminescens or the PpyRE-TS and PpyGR-TS firefly luciferase mutants of Photinus pyralis (red and green thermostable P. pyralis luciferase mutants, respectively) have been engineered and used to monitor intestinal colonization in the streptomycin-treated mouse model. There was excellent correlation between the bioluminescence signal measured in the feces (R 2 ‫؍‬ 0.98) or transcutaneously in the abdominal region of whole animals (R 2 ‫؍‬ 0.99) and the CFU counts in the feces of bacteria harboring the luxABCDE operon. Stability in vivo of the bioluminescence signal was achieved by constructing plasmid pAT881(pGB2⍀P ami luxABCDE), which allowed long-term monitoring of intestinal colonization without the need for antibiotic selection for plasmid maintenance. Levels of intestinal colonization by various strains of E. coli could be compared directly by simple recording of the bioluminescence signal in living animals. The difference in spectra of light emission of the PpyRE-TS and PpyGR-TS firefly luciferase mutants and dual bioluminescence detection allowed direct in vitro and in vivo quantification of two bacterial populations by measurement of red and green emitted signals and thus monitoring of the two populations simultaneously. This system offers a simple and direct method to study in vitro and in vivo competition between mutants and the parental strain. BLI is a useful tool to study intestinal colonization.Among the wide variety of bacteria that colonize the gastrointestinal tracts of mammals, Escherichia coli is the most abundant facultative anaerobe of the human intestinal microflora. Aside from being part of the normal flora, E. coli is also a versatile organism capable of causing a variety of intestinal and extraintestinal diseases (18). The mechanisms that allow commensal E. coli to colonize the intestine and survive successfully in this niche remain poorly characterized. Conventional mice display natural resistance to colonization by commensal E. coli, but oral administration of streptomycin, which alters the intestinal microflora, allows colonization of the mouse large intestine by this species (25). The streptomycin-treated mouse model has been used extensively to study the factors of gramnegative bacteria implicated in the intestinal colonization process. However, this model is limited to the viable plate counts of bacteria in the feces and misses some critical information, such as the kinetics of colonization, the fate of the bacterial cells across the digestive tract, and the site of colonization. A better understanding of colonization would be facilitated by direct in vivo follow-up of this process.Biolumine...

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A redshifted codon-optimized firefly luciferase is a sensitive reporter for bioluminescence imaging

Cited by 48 publications

References 31 publications

In Vivo Bioluminescence Imaging of Murine Xenograft Cancer Models with a Red-shifted Thermostable Luciferase

In Vivo Bioluminescence Imaging of Murine Xenograft Cancer Models with a Red-shifted Thermostable Luciferase

In vivo bioluminescence for tracking cell fate and function

In Vivo Bioluminescence Imaging for the Study of Intestinal Colonization by Escherichia coli in Mice

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