Substrate-mediated gene delivery is a novel delivery system that maintains elevated DNA concentrations in the cell microenvironment. It functions by self-assembling DNA with cationic polymers or lipids, resulting in positively charged complexes which can interact with a cell-adhesive substrate, which can in turn enhance the transfection efficiency of cells cultured on the substrate. I am using self-assembled monolayers (SAMs) of
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alkanethiols on gold and spotting technology to investigate
the influence of various surface properties on DNA complex binding
and surface-mediated transfection. Using
various alkanethiols which vary in their terminal functional group,
surface properties can be varied and correlated to the ability of
DNA complexes to adsorb and subsequently desorb from the surface
and transfect cells. I have found that charged, hydrophilic surfaces
are able to support the highest transfection efficiencies using model
cell lines, as well as breast cancer cells. Using my controlled surface
chemistries and microarray patterning strategies, combined with plasmids
containing reporter genes fused with promoters of genes important
to estrogen receptor signaling in breast cancer, I hope to correlate
induction levels of these promoters, in response to various coactivators
or cosupressors, to tumor state. Initially, I have proven that substrate-mediated
delivery is capable of reporting on induction of these various
pathways. Eventually, this system will then be used to characterize
the expression of several related genes simultaneously on a single
surface and correlate expression to disease diagnosis and treatment. In
addition to creating this transfected cell array to report on the
activation of the estrogen receptor in breast cancer, I am also
utilizing the substrate mediated delivery system as a method to
transfect mouse embryonic stem cells. Angela's publications:
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