Predictive modeling of non-viral gene transfer

Authors

  • Gerlinde Schwake,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
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    • Gerlinde Schwake, Simon Youssef, and Jan-Timm Kuhr contributed equally to this work.

  • Simon Youssef,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
    2. Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 München, Germany
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    • Gerlinde Schwake, Simon Youssef, and Jan-Timm Kuhr contributed equally to this work.

  • Jan-Timm Kuhr,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
    2. Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 München, Germany
    3. Arnold Sommerfeld Center for Theoretical Physics, Geschwister-Scholl-Platz 1, D-80539 München, Germany
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    • Gerlinde Schwake, Simon Youssef, and Jan-Timm Kuhr contributed equally to this work.

  • Sebastian Gude,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
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  • Maria Pamela David,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
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  • Eduardo Mendoza,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
    2. Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 München, Germany
    3. Institute of Mathematics, University of the Philippines, Diliman, Quezon City 1101, Philippines
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  • Erwin Frey,

    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
    2. Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 München, Germany
    3. Arnold Sommerfeld Center for Theoretical Physics, Geschwister-Scholl-Platz 1, D-80539 München, Germany
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  • Joachim O. Rädler

    Corresponding author
    1. Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182
    2. Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 München, Germany
    • Fakultät für Physik, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, D-80539 München, Germany; telephone: +49-(0)89-2180-2438; fax: +49-(0)89-2180-3182.
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Abstract

In non-viral gene delivery, the variance of transgenic expression stems from the low number of plasmids successfully transferred. Here, we experimentally determine Lipofectamine- and PEI-mediated exogenous gene expression distributions from single cell time-lapse analysis. Broad Poisson-like distributions of steady state expression are observed for both transfection agents, when used with synchronized cell lines. At the same time, co-transfection analysis with YFP- and CFP-coding plasmids shows that multiple plasmids are simultaneously expressed, suggesting that plasmids are delivered in correlated units (complexes). We present a mathematical model of transfection, where a stochastic, two-step process is assumed, with the first being the low-probability entry step of complexes into the nucleus, followed by the subsequent release and activation of a small number of plasmids from a delivered complex. This conceptually simple model consistently predicts the observed fraction of transfected cells, the cotransfection ratio and the expression level distribution. It yields the number of efficient plasmids per complex and elucidates the origin of the associated noise, consequently providing a platform for evaluating and improving non-viral vectors. Biotechnol. Bioeng. 2010. 105: 805–813. © 2009 Wiley Periodicals, Inc.

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