Predictive Modeling of Non-Viral Gene Transfer - Quantitative Biology > Quantitative MethodsReport as inadecuate

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Abstract: In non-viral gene delivery, the variance of transgenic expression stems fromthe low number of plasmids successfully transferred. Here, we experimentallydetermine Lipofectamine- and PEI-mediated exogenous gene expressiondistributions from single cell time-lapse analysis. Broad Poisson-likedistributions of steady state expression are observed for both transfectionagents, when used with synchronized cell lines. At the same time,co-transfection analysis with YFP- and CFP-coding plasmids shows that multipleplasmids are simultaneously expressed, suggesting that plasmids are deliveredin correlated units complexes. We present a mathematical model oftransfection, where a stochastic, two-step process is assumed, with the firstbeing the low-probability entry step of complexes into the nucleus, followed bythe subsequent release and activation of a small number of plasmids from adelivered complex. This conceptually simple model consistently predicts theobserved fraction of transfected cells, the cotransfection ratio and theexpression level distribution. It yields the number of efficient plasmids percomplex and elucidates the origin of the associated noise, consequentlyproviding a platform for evaluating and improving non-viral vectors.

Author: Gerlinde Schwake, Simon Youssef, Jan-Timm Kuhr, Sebastian Gude, Maria Pamela David, Eduardo Mendoza, Erwin Frey, Joachim O. Rädl


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