2017
9TH NATIONAL CONGRESS WITH INTERNATIONAL PARTICIPATION AND 35TH ANNUAL SCIENTIFIC SESSION OF ROMANIAN SOCIETY FOR CELL BIOLOGY, June 7-11, Iasi
Daniela Rebleanu, Cristina Ana Constantinescu, Monica Tucureanu, Geanina Voicu, Elena Butoi, Agneta Simionescu, Ileana Manduteanu, Manuela Calin
Institute of Cellular Biology and Pathology “Nicolae Simionescu” of Romanian Academy, Bucharest, Romania
Introduction. RNA-interference (RNAi) is a strong therapeutic tool to silence the expression of an undesirable gene. Short hairpin (sh) RNA is an advantageous mediator of RNAi due to its relatively low rate of degradation and is synthesized within the cell after transfection with a plasmid shRNA. The aim of this study was to obtain lipid-based transfection nanocarriers to function as efficient vectors for RNAi in valvular cells.
Methods. Different formulations of cationic liposomes with different lipid compositions were designed and prepared in order to find the optimal composition for efficient delivery of shRNA to cells. Cationic liposomes were complexed with a plasmid shRNA at various electric charge ratios (R) +/- and characterized by determining the distribution of dimensions for liposomes and liposomes/shRNA plasmid lipoplexes (by dynamic light scattering) and the capacity of liposomes to protect shRNA (by agarose gel retardation assay).
Results. 1) the size of liposomes was ~200 nm, whereas the size of lipoplexes varies as function of R (between 100 and 2500 nm) and 2) the migration of plasmid shRNA in agarose gel is completely blocked for charge ratio +/- starting with 4.
Acknowledgements. This work is supported by the Competitiveness Operational Programme 2014-2020, Priority Axis1/Action 1.1.4/, Financing Contract no.115/13.09.2016/ MySMIS:104362.
