1 SCIENTIFIC REPORTS | (2020) 10:2121 | https://doi.org/10.1038/s41598-020-59075-3 www.nature.com/scientificreports Generation of light-producing somatic-transgenic mice using adeno-associated virus vectors Rajvinder Karda 1 , Ahad A. Rahim 2 , Andrew M. S. Wong 3 , Natalie Suf 1 , Juan Antinao Diaz 1 , Dany P. Perocheau 1 , Maha Tijani 1 , Joanne Ng 1 , Julien Baruteau 1 , Nuria Palomar Martin 4 , Michael Hughes 2 , Juliette M. K. M. Delhove 5 , John R. Counsell 6,7 , Jonathan D. Cooper 3,8 , Els Henckaerts 4,9 , Tristan R. Mckay 10 , Suzanne M. K. Buckley 1* & Simon N. Waddington 1,11 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 frst 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 frefy 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 infammation. This technology complements existing germline-transgenic models and may be applicable to other rodent disease models. Germline light-producing transgenic mice where luciferase expression is controlled by an endogenous promoter, a surrogate promoter or by a minimal promoter downstream of tandem, synthetic, transcription factor binding elements, are used to provide an in vivo readout of physiological and pathological processes 1,2 . One of the advan- tages of this technology is that every cell will contain a copy of the luciferase transgene and therefore provide a whole-body transgene expression profle under the control of a specifc promoter of choice. However, producing germline transgenics requires frequent backcrossing and therefore becomes a time-consuming and costly process, using many rodents. We have previously developed a novel technology which allows the generation of light-producing somatic transgenic rodents, using lentiviral vectors as a proof-of-concept system 3 and have validated this technology both in vitro 4,5 and in vivo 1,6 . We have also demonstrated that signalling pathways in diseased organs can be moni- tored specifcally, continually and in a non-invasive manner 1,2 . Exploiting the immune tolerance of neonatal mice 1 Gene Transfer Technology Group, Institute for Women’s Health, University College London, London, UK. 2 UCL School of Pharmacy, University College London, London, UK. 3 Institute of Psychiatry, Psychology & Neuroscience, King’s College London, London, UK. 4 Department of Infectious Diseases, School of Immunology and Microbial Sciences, King’s College London, London, UK. 5 Robinson Research Institute, University of Adelaide, Adelaide, Australia. 6 Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Programme, UCL Great Ormond Street Institute of Child Health, London, UK. 7 NIHR Great Ormond Street Hospital Biomedical Research Centre, London, UK. 8 Department of Pediatrics, Washington University in St Louis, St Louis, MO, USA. 9 Laboratory of Viral Cell Signalling and Therapeutics, Department of Cellular and Molecular Medicine and Department of Microbiology, Immunology and Transplantation, KU Leuven, 3000, Leuven, Belgium. 10 Centre for Biomedicine, Manchester Metropolitan University, Manchester, UK. 11 Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa. *email: suzy.buckley@ucl.ac.uk OPEN