Pannier Lab
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Understanding Cell Response to Nonviral Vectors Through Cell Priming

 
Sarah Plautz, Andrew Hamann, and Albert Nguyen

The long-term goal of this project is to determine and dynamically model the “biology of transfection” in order to exploit the mechanisms that render cells responsive to DNA transfer for improved transfection. The untransfected cells, those treated with DNA complexes but not displaying transgene expression, prevent therapeutic application of nonviral DNA delivery. Attempts in the field to improve transfection have included DNA carrier modifications to overcome extra- and intra-cellular barriers. More recently, microarray analyses have been used to identify the pharmacogenomic responses to nonviral carriers as strategies for improving carrier design and transfection. Even with these efforts the molecular mechanisms that facilitate successful DNA transfer remain unclear and the efficiency of nonviral gene delivery is still below desired levels. Therefore, there is a need to identify the endogenous mechanisms that facilitate DNA transfer for improved carrier design and to prepare the cell for optimal DNA delivery.

The central hypothesis for this research is that endogenous mechanisms characteristic of transfected cells facilitate DNA transfer while untransfected cells fail to express these mechanisms or are untimely in their expression. Using microarray analysis, we have identified genes that are up- and down-regulated during the DNA transfer process at 2h, 8h, 16h, and 24h time points. Furthermore, priming cells by perturbing these gene pathways using pharmacologic agents resulted in increased transfection and identified specific molecules that affect DNA transfer. With these results, future work includes cell priming for efficient DNA transfer or engineering strategies to modify the DNA carrier to include targeting moieties that associate with key molecules of DNA transfer.