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Generation of light-producing somatic-transgenic mice using adeno-associated virus vectors

Karda, R and Rahim, AA and Wong, AMS and Suff, N and Diaz, JA and Perocheau, DP and Tijani, M and Ng, J and Baruteau, J and Martin, NP and Hughes, M and Delhove, JMKM and Counsell, JR and Cooper, JD and Henckaerts, E and Mckay, TR and Buckley, SMK and Waddington, SN (2020) Generation of light-producing somatic-transgenic mice using adeno-associated virus vectors. Scientific Reports, 10 (1). ISSN 2045-2322


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© 2020, The Author(s). We have previously designed a library of lentiviral vectors to generate somatic-transgenic rodents to monitor signalling pathways in diseased organs using whole-body bioluminescence imaging, in conscious, freely moving rodents. We have now expanded this technology to adeno-associated viral vectors. We first explored bio-distribution by assessing GFP expression after neonatal intravenous delivery of AAV8. We observed widespread gene expression in, central and peripheral nervous system, liver, kidney and skeletal muscle. Next, we selected a constitutive SFFV promoter and NFκB binding sequence for bioluminescence and biosensor evaluation. An intravenous injection of AAV8 containing firefly luciferase and eGFP under transcriptional control of either element resulted in strong and persistent widespread luciferase expression. A single dose of LPS-induced a 10-fold increase in luciferase expression in AAV8-NFκB mice and immunohistochemistry revealed GFP expression in cells of astrocytic and neuronal morphology. Importantly, whole-body bioluminescence persisted up to 240 days. We have validated a novel biosensor technology in an AAV system by using an NFκB response element and revealed its potential to monitor signalling pathway in a non-invasive manner in a model of LPS-induced inflammation. This technology complements existing germline-transgenic models and may be applicable to other rodent disease models.

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