Development of an inducible mouse model of iRFP713 to track recombinase activity and tumour development in vivo

Scientific Reports
Andreas K HockKaren H Vousden

Abstract

While the use of bioluminescent proteins for molecular imaging is a powerful technology to further our understanding of complex processes, fluorescent labeling with visible light fluorescent proteins such as GFP and RFP suffers from poor tissue penetration and high background autofluorescence. To overcome these limitations, we generated an inducible knock-in mouse model of iRFP713. This model was used to assess Cre activity in a Rosa Cre-ER background and quantify Cre activity upon different tamoxifen treatments in several organs. We also show that iRFP can be readily detected in 3D organoid cultures, FACS analysis and in vivo tumour models. Taken together we demonstrate that iRFP713 is a progressive step in in vivo imaging and analysis that widens the optical imaging window to the near-infrared spectrum, thereby allowing deeper tissue penetration, quicker image acquisition without the need to inject substrates and a better signal to background ratio in genetically engineered mouse models (GEMMs).

References

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Jun 19, 2013·Nature Methods·Daria M Shcherbakova, Vladislav V Verkhusha
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Nov 12, 2016·Scientific Reports·Konstantin A RumyantsevVladislav V Verkhusha

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Citations

Aug 5, 2017·International Journal of Molecular Sciences·Olena S OliinykVladislav V Verkhusha
Jun 22, 2018·Science Translational Medicine·Björn KruspigDaniel J Murphy
Sep 13, 2017·Scientific Reports·Palani SharmiladeviAgnishwar Girigoswami
Jun 20, 2019·Biochemistry. Biokhimii︠a︡·M M KarasevV V Verkhusha
Dec 14, 2018·Cancer & Metabolism·Timothy J HumptonKaren H Vousden
Jul 26, 2018·Trends in Biotechnology·Daria M ShcherbakovaVladislav V Verkhusha

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Methods Mentioned

BETA
xenograft
FACS
imaging techniques
fluorescence imaging
transgenic

Software Mentioned

Image Studio
BD
Prism Graph Pad
FACSdiva
Volocity
Image

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