title: In vivo bioimaging with tissue-specific transcription factor activated luciferase reporters.
abstract: The application of transcription factor activated luciferase reporter cassettes in vitro is widespread but potential for in vivo application has not yet been realized. Bioluminescence imaging enables non-invasive tracking of gene expression in transfected tissues of living rodents. However the mature immune response limits luciferase expression when delivered in adulthood. We present a novel approach of tissue-targeted delivery of transcription factor activated luciferase reporter lentiviruses to neonatal rodents as an alternative to the existing technology of generating germline transgenic light producing rodents. At this age, neonates acquire immune tolerance to the conditionally responsive luciferase reporter. This simple and transferrable procedure permits surrogate quantitation of transcription factor activity over the lifetime of the animal. We show principal efficacy by temporally quantifying NFκB activity in the brain, liver and lungs of somatotransgenic reporter mice subjected to lipopolysaccharide (LPS)-induced inflammation. This response is ablated in Tlr4(-/-) mice or when co-administered with the anti-inflammatory glucocorticoid analogue dexamethasone. Furthermore, we show the malleability of this technology by quantifying NFκB-mediated luciferase expression in outbred rats. Finally, we use somatotransgenic bioimaging to longitudinally quantify LPS- and ActivinA-induced upregulation of liver specific glucocorticoid receptor and Smad2/3 reporter constructs in somatotransgenic mice, respectively.
authors.affiliation: Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
authors.forename: Suzanne M K
authors.initials: SM
authors.lastname: Buckley
authors.affiliation: 1] Stem Cell Group, Cardiovascular &Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK [2] Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.
authors.forename: Juliette M K M
authors.initials: JM
authors.lastname: Delhove
authors.affiliation: Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
authors.forename: Dany P
authors.initials: DP
authors.lastname: Perocheau
authors.affiliation: 1] Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK [2] Faculty of Medicine, Department of Surgery &Cancer, Imperial College, London, UK.
authors.forename: Rajvinder
authors.initials: R
authors.lastname: Karda
authors.affiliation: Department of Pharmacology, School of Pharmacy, University College London, 29-39 Brunswick Square, London WC1N 1AX, UK.
authors.forename: Ahad A
authors.initials: AA
authors.lastname: Rahim
authors.affiliation: Wolfson Institute for Gene Therapy, Molecular and Cellular Immunology, Institute of Child Health, University College London, London WC1N 1EH, UK.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Howe
authors.affiliation: Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
authors.forename: Natalie J
authors.initials: NJ
authors.lastname: Ward
authors.affiliation: Faculty of Medicine, National Heart &Lung Institute, Imperial College, London, UK.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Birrell
authors.affiliation: Faculty of Medicine, National Heart &Lung Institute, Imperial College, London, UK.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Belvisi
authors.affiliation: Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.
authors.forename: Patrick
authors.initials: P
authors.lastname: Arbuthnot
authors.affiliation: Faculty of Medicine, Department of Surgery &Cancer, Imperial College, London, UK.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Johnson
authors.affiliation: 1] Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK [2] Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.
authors.forename: Simon N
authors.initials: SN
authors.lastname: Waddington
authors.affiliation: Stem Cell Group, Cardiovascular &Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK.
authors.forename: Tristan R
authors.initials: TR
authors.lastname: McKay
mesh_terms: D000818:Animals; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D041681:NIH 3T3 Cells; D009928:Organ Specificity; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000502:physiology*; D015533:Transcriptional Activation / Q000276:immunology*
chemical_list: D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly