NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice-A Novel in Vivo Imaging Platform.

Autor:	Gaspar N; Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands.; Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands.; Percuros B.V., 2333 CL Leiden, The Netherlands., Zambito G; Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands.; Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands.; Medres Medical Research GMBH, 50931 Cologne, Germany., Dautzenberg IJC; Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands., Cramer SJ; Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands., Hoeben RC; Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands., Lowik C; Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands.; Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands.; Department of Oncology CHUV, UNIL and Ludwig Cancer Center, 1011 Lausanne, Switzerland., Walker JR; Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA., Kirkland TA; Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA., Smith TP; Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA., van Weerden WM; Erasmus Medical Center, Department of Urology, 3015 GD Rotterdam, The Netherlands., de Vrij J; Erasmus Medical Center, Department of Neurosurgery, 3015 GD Rotterdam, The Netherlands., Mezzanotte L; Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands.; Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands.
Jazyk:	angličtina
Zdroj:	International journal of molecular sciences [Int J Mol Sci] 2020 Aug 15; Vol. 21 (16). Date of Electronic Publication: 2020 Aug 15.
DOI:	10.3390/ijms21165863
Abstrakt:	Reporter genes are used to visualize intracellular biological phenomena, including viral infection. Here we demonstrate bioluminescent imaging of viral infection using the NanoBiT system in combination with intraperitoneal injection of a furimazine analogue, hydrofurimazine. This recently developed substrate has enhanced aqueous solubility allowing delivery of higher doses for in vivo imaging. The small high-affinity peptide tag (HiBiT), which is only 11 amino-acids in length, was engineered into a clinically used oncolytic adenovirus, and the complementary large protein (LgBiT) was constitutively expressed in tumor cells. Infection of the LgBiT expressing cells with the HiBiT oncolytic virus will reconstitute NanoLuc in the cytosol of the cell, providing strong bioluminescence upon treatment with substrate. This new bioluminescent system served as an early stage quantitative viral transduction reporter in vitro and also in vivo in mice, for longitudinal monitoring of oncolytic viral persistence in infected tumor cells. This platform provides novel opportunities for studying the biology of viruses in animal models.
Databáze:	MEDLINE
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