title: Mapping of bionic array electric field focusing in plasmid DNA-based gene electrotransfer.
abstract: Molecular medicine through gene therapy is challenged to achieve targeted action. This is now possible utilizing bionic electrode arrays for focal delivery of naked (plasmid) DNA via gene electrotransfer. Here, we establish the properties of array-based electroporation affecting targeted gene delivery. An array with eight 300 μm platinum ring electrodes configured as a cochlear implant bionic interface was used to transduce HEK293 cell monolayers with a plasmid-DNA green fluorescent protein (GFP) reporter gene construct. Electroporation parameters were pulse intensity, number, duration, separation and electrode configuration. The latter determined the shape of the electric fields, which were mapped using a voltage probe. Electrode array-based electroporation was found to require ~100 × lower applied voltages for cell transduction than conventional electroporation. This was found to be due to compression of the field lines orthogonal to the array. A circular area of GFP-positive cells was created when the electrodes were ganged together as four adjacent anodes and four cathodes, whereas alternating electrode polarity created a linear area of GFP-positive cells. The refinement of gene delivery parameters was validated in vivo in the guinea pig cochlea. These findings have significant clinical ramifications, where spatiotemporal control of gene expression can be predicted by manipulation of the electric field via current steering at a cellular level.
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.|School of Science and Health, Western Sydney University, Sydney, New South Wales, Australia.
authors.forename: C J
authors.initials: CJ
authors.lastname: Browne
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: J L
authors.initials: JL
authors.lastname: Pinyon
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: D M
authors.initials: DM
authors.lastname: Housley
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: E N
authors.initials: EN
authors.lastname: Crawford
authors.affiliation: Graduate School of Biomedical Engineering, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: N H
authors.initials: NH
authors.lastname: Lovell
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: M
authors.identifier: http://orcid.org/0000-0003-0574-612X
authors.initials: M
authors.lastname: Klugmann
authors.affiliation: Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Australia, Sydney, New South Wales, Australia.
authors.forename: G D
authors.initials: GD
authors.lastname: Housley
mesh_terms: D000818:Animals; D001701:Bionics / Q000295:instrumentation / Q000379:methods; D004566:Electrodes; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006168:Guinea Pigs; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics
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