Predictive modeling of non-viral gene transfer

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 distrib...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2010-03, Vol.105 (4), p.805-813
Main Authors: Schwake, Gerlinde, Youssef, Simon, Kuhr, Jan-Timm, Gude, Sebastian, David, Maria Pamela, Mendoza, Eduardo, Frey, Erwin, Rädler, Joachim O
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Cells
Epithelial Cells - cytology
Epithelial Cells - metabolism
Fundamental and applied biological sciences. Psychology
Gene expression
gene transfer
Green Fluorescent Proteins - analysis
Green Fluorescent Proteins - genetics
Humans
Imines - administration & dosage
Lipids - administration & dosage
mathematical modeling
Mathematical models
Models, Genetic
Plasmids
Plasmids - administration & dosage
Polyethylenes - administration & dosage
single cell
Transfection
transfection/gene expression
Viruses
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Summary: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.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.22604