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• W2317177124 abstract "Event Abstract Back to Event Inhibition of interferon α expression with small interference RNA increases adenoviruses-GFP transduction and transgene expression in Huh7 cells Ana A. Sobrevilla-Navarro1*, Ana S. Sandoval-Rodríguez1, Jesús García-Bañuelos1, Pedro Sánchez-Hernández2, Luis D. Hernández-Ortega1, Jorge Gaona-Bernal3, Juan Armendáriz-Borunda1 and Adriana M Salazar-Montes1 1 University of Guadalajara, Department of Molecular Biology and Genomics, Mexico 2 University of Guadalajara , Laboratory of Immunology, Mexico 3 National Autonomous University of Mexico, Department of Microbiology and Parasitology, Mexico Introduction: Recombinant adenoviruses (rAd) are the most commonly vectors used in clinical trials of gene therapy. Systemic administration of rAd present high tropism for liver. However, host´s immune response against rAd orchestrated by Interferons type 1 (α and beta) limits the therapeutic gene expression and prevents new administrations. Aim: To evaluate the effect of IFNα inhibition by a small interfering RNA (siRNA) on rAd-GFP transduction and transgene expression in Huh7 cell line. Methods: Huh7 cells are cultured in DMEM, 5% FBS at 37 °C and 5%CO2 and then transfected with 70 nM of siRNA-IFNα. 6h later culture was exposed to 1.0 x 109 vp/ml of rAd-GFP for 24 hrs. Expression of IFNα1 and TNF-α were determined by qRT-PCR . Cell transduction was analyzed by fluorescent analysis cell sorting (FACS) and qPCR. GFP expression was determined by western blot. Results: 70 nM of siRNA-IFNα1 inhibited 96% of IFNα1 gene expression (p<0.001) and 65% of TNF-α (p<0.05) respect to control siRNA-irrelevant. Transduction and transgen expression were increased in cells treated with siRNA-IFNα1 compared to control. Conclusions: Inhibition of IFNα by siRNA-IFNα1 permits a higher transgene expression (GFP) indicating the crucial role of IFNα on adenoviruses elimination in transducted cells. This strategy could be useful in clinical trials directed to liver diseases, where adenoviruses are used as vectors for therapeutic genes; achieving an increased expression of these genes leading to better results in liver diseases resolution. References  Wiley, J. (2012). The journal of Gene Medicine.  Arnberg, N. (2009). Adenovirus receptors: implications for tropism, treatment and targeting. RevMedVirol 19, 165-178.  Elbashir, S.M., Martinez, J., Patkaniowska, A., Lendeckel, W., and Tuschl, T. (2001). Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. EMBO J 20, 6877-6888.  Thaci, B., Ulasov, I.V., Wainwright, D.A., and Lesniak, M.S. (2011). The challenge for gene therapy: innate immune response to adenoviruses. Oncotarget 2, 113-121.  Zhu, J., Huang, X., and Yang, Y. (2008). A critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo. MolTher 16, 1300-130 Keywords: Adenoviruses, IFNα, innate immune response, siRNA, Huh7 Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Innate immunity Citation: Sobrevilla-Navarro AA, Sandoval-Rodríguez A, García-Bañuelos J, Sánchez-Hernández P, Hernández-Ortega L, Gaona-Bernal J, Armendáriz-Borunda J and Salazar-Montes A (2013). Inhibition of interferon α expression with small interference RNA increases adenoviruses-GFP transduction and transgene expression in Huh7 cells. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00990 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 26 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Ana A Sobrevilla-Navarro, University of Guadalajara, Department of Molecular Biology and Genomics, Guadalajara, 44340, Mexico, alondra280381@hotmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Ana A Sobrevilla-Navarro Ana S. Sandoval-Rodríguez Jesús García-Bañuelos Pedro Sánchez-Hernández Luis D. Hernández-Ortega Jorge Gaona-Bernal Juan Armendáriz-Borunda Adriana M Salazar-Montes Google Ana A Sobrevilla-Navarro Ana S. Sandoval-Rodríguez Jesús García-Bañuelos Pedro Sánchez-Hernández Luis D. Hernández-Ortega Jorge Gaona-Bernal Juan Armendáriz-Borunda Adriana M Salazar-Montes Google Scholar Ana A Sobrevilla-Navarro Ana S. Sandoval-Rodríguez Jesús García-Bañuelos Pedro Sánchez-Hernández Luis D. Hernández-Ortega Jorge Gaona-Bernal Juan Armendáriz-Borunda Adriana M Salazar-Montes PubMed Ana A Sobrevilla-Navarro Ana S. Sandoval-Rodríguez Jesús García-Bañuelos Pedro Sánchez-Hernández Luis D. Hernández-Ortega Jorge Gaona-Bernal Juan Armendáriz-Borunda Adriana M Salazar-Montes Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page." @default.
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• W2317177124 title "Inhibition of interferon a expression with small interference RNA increases adenoviruses-GFP transduction and transgene expression in Huh7 cells" @default.
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