Berlin 2018 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 9: Postersession I
BP 9.40: Poster
Montag, 12. März 2018, 17:30–19:30, Poster A
Time distribution of mRNA delivery mediated by lipoplexes determined from single cell expression onsets — •Anita Reiser1,2, Neha Mehrotra1, Rafal Krzyszton1,2, Daniel Woschée1, Helmut H. Strey3, and Joachim O. Rädler1,2 — 1Faculty of Physics and Center for NanoScience, Ludwig-Maximilians-University, Munich, Germany — 2Graduate School of Quantitative Biosciences (QBM), Ludwig-Maximilians-University, Munich, Germany — 3Department of Biomedical Engineering and Laufer Center for Quantitative Biology, Stony Brook University, Stony Brook, NY, USA
Nucleic acid (NA) based therapies require efficient delivery systems. New Methods are required to assess uptake and release kinetics in living cells in order to improve the internalization of NA carriers. Here we employ automated time-lapse microscopy to study the delivery-time distribution of lipid based mRNA vectors. We determine the protein expression onset-times after transfection from single cell time courses of eGFP reporter fluorescence. We show how maximum likelihood fitting yields best estimates of individual onset times from hundreds of individual time courses in a robust and automated manner. The distribution of delivery dwell-times is found to be log-normal distributed in all cases with the mean depending on the transfection agent. For cationic lipid formulation containing various amounts of cholesterol the delivery timing correlates with efficiency. The profile of delivery timing might be a valuable indicator for the development of nanocarriers with improved uptake activity and endosomal release kinetics.