Istry of HPV E7 Proteins Biological Activity Science ICT (2017M3A9G8083382).ISEV 2018 abstract bookPS02.Novel antibody-mediated drug delivery technique for targeting exosomal microRNA Asako Yamayoshi1; Ryo Konishi2; Akio Kobori2; Naoto Yamashita3; Eishi Ashihara3; Akira Murakami3; Hiroshi Sugiyama4 The Hakubi Cemter for Sophisticated Study, Kyoto Univiersity, Kyoto, Japan; 2Department of Biomolecular Engineering, Kyoto Institute of Technologies, Kyoto, Japan; 3Department of Clinical and Translational Physiology, Kyoto Pharmaceutical University, Kyoto, Japan; 4Graduate School of Science, Kyoto University, Kyoto, JapanBackground: Recently, microRNAs (miRNAs) happen to be identified in exosomes, which is often taken up by neighbouring or distant cells. It has also been reported that such miRNAs (exosomal-miRNAs) regulates gene expression in the recipient cells. In accordance with current reports, the aberrant expression of miRNAs is connected with most pathological disease processes, such as carcinogenesis. Hence, exosomalmiRNAs are deemed as considerable therapeutic targets for cancer therapy. Nonetheless, there is certainly no report to regulate the function of miRNAs in exosomes. In this study, we attempt to create novel drug delivery technique applying anti-exosome antibody ligonucleotide conjugates (ExomiR-Tracker) for functional inhibition of exosomal-miRNAs. Solutions: Cellular uptakes and localization of ExomiR-Trackers were evaluated by confocal microscopy. We evaluate the inhibitory effects of ExomiR-Tracker in accordance with earlier report (Ariyoshi et al., Bioconj Chem. 2015). Cystatin-1 Proteins supplier Benefits: Initially, we evaluated cellular uptakes and localization of anti-miRconjugated antibody by confocal microscopy. In this experiment, Alexa647-labelled anti-miR and cationized anti-exosome antibody had been employed for ExomiR-Tracker. Fluorescence signals have been successfully observed inside HeLa cells. In contrast, inside the case of control molecules, no fluorescent signals were observed. Next, we evaluated inhibitory effects of ExomiR-Tracker against miRNA functions. It was identified that luminescence intensity of ExomiR-Trackertreated cells was recovered in comparison with the case of manage ExomiRTracker. This outcome suggests that ExomiR-Tracker effectively inhibit the function of miR-Luc in HeLa cells. We also confirmed these effects in vivo. Summary/Conclusion: We successfully demonstrated that ExomiRTracker is incorporated in to the recipient cells and inhibits miRNA function in vitro and in vivo. Towards the most effective of our information, this really is the initial instance of regulating exosomal-miRNA within the recipient cells. Funding: This study was partly supported by a Grant-in-Aid for Scientific Study in the Ministry of Education, Science, Sports and Culture of JapanGrant No. 15K05564and JST, PRESTO (Grant No. JPMJPR178A), Japan.new therapeutic paradigms in illness. However, deliverability and stability of RNA-based drugs is still restricted, that is mostly resulting from the lack of proper delivery systems. Current studies have shown that EVs are organic carriers of miRNA and this intrinsic home could possibly be exploited as a gene delivery method. Existing approaches for loading of EVs with RNA are in vitro electroporation, transfection, co-incubation, co-expression of target RNA and zipcoding but all of these suffer from poor efficiency. Our study aims to work with RNA binding proteins (RBP) fused to EV marker proteins for in vitro loading of EVs with cargo RNA tagged using the cognate RNA recognition components. Techniques: A RNA library of target RNA fused to a spec.