{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T03:05:51Z","timestamp":1775012751054,"version":"3.50.1"},"reference-count":25,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2020,8,15]],"date-time":"2020-08-15T00:00:00Z","timestamp":1597449600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Commission","doi-asserted-by":"publisher","award":["777682"],"award-info":[{"award-number":["777682"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000780","name":"European Commission","doi-asserted-by":"publisher","award":["675743"],"award-info":[{"award-number":["675743"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/ijms21165863","type":"journal-article","created":{"date-parts":[[2020,8,17]],"date-time":"2020-08-17T04:35:51Z","timestamp":1597638951000},"page":"5863","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice\u2014A Novel in Vivo Imaging Platform"],"prefix":"10.3390","volume":"21","author":[{"given":"Natasa","family":"Gaspar","sequence":"first","affiliation":[{"name":"Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands"},{"name":"Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands"},{"name":"Percuros B.V., 2333 CL Leiden, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giorgia","family":"Zambito","sequence":"additional","affiliation":[{"name":"Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands"},{"name":"Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands"},{"name":"Medres Medical Research GMBH, 50931 Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iris J. C.","family":"Dautzenberg","sequence":"additional","affiliation":[{"name":"Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steve J.","family":"Cramer","sequence":"additional","affiliation":[{"name":"Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rob C.","family":"Hoeben","sequence":"additional","affiliation":[{"name":"Leiden University Medical Center, Department of Cell and Chemical Biology, 2333 Leiden, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0859-5785","authenticated-orcid":false,"given":"Clemens","family":"Lowik","sequence":"additional","affiliation":[{"name":"Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands"},{"name":"Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands"},{"name":"Department of Oncology CHUV, UNIL and Ludwig Cancer Center, 1011 Lausanne, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joel R.","family":"Walker","sequence":"additional","affiliation":[{"name":"Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0324-4804","authenticated-orcid":false,"given":"Thomas A.","family":"Kirkland","sequence":"additional","affiliation":[{"name":"Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1381-9360","authenticated-orcid":false,"given":"Thomas P.","family":"Smith","sequence":"additional","affiliation":[{"name":"Promega Biosciences L.L.C., San Luis Obispo, CA 93401, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9443-8377","authenticated-orcid":false,"given":"Wytske M.","family":"van Weerden","sequence":"additional","affiliation":[{"name":"Erasmus Medical Center, Department of Urology, 3015 GD Rotterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeroen","family":"de Vrij","sequence":"additional","affiliation":[{"name":"Erasmus Medical Center, Department of Neurosurgery, 3015 GD Rotterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Mezzanotte","sequence":"additional","affiliation":[{"name":"Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands"},{"name":"Erasmus Medical Center, Department of Molecular Genetics, 3015 GD Rotterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.2147\/OV.S137159","article-title":"The importance of imaging strategies for pre-clinical and clinical in vivo distribution of oncolytic viruses","volume":"7","author":"Pelin","year":"2018","journal-title":"Oncolytic Virother."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"169","DOI":"10.2217\/fvl.14.96","article-title":"A bright future for bioluminescent imaging in viral research","volume":"10","author":"Coleman","year":"2015","journal-title":"Future Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"14610","DOI":"10.1073\/pnas.1212379109","article-title":"Dengue reporter viruses reveal viral dynamics in interferon receptor-deficient mice and sensitivity to interferon effectors in vitro","volume":"109","author":"Schoggins","year":"2012","journal-title":"Proc. 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