Fabrication of an Innovative Nanopore Microneedle for Controlled Electroporation and Gene Transfection

A significant challenge for in vivo genetic engineering is the transmembrane transport of relatively large genetic payloads. Larger molecules and transcription plasmids typically require viral vectors or permeabilizing agents. which can be challenging to engineer and may have deleterious systemic effects. As an alternative approach. electroporation uses ultrashort electrical pulses to transport large molecules across cell membranes. This approach is advantageous in applications such as delivery of DNA vaccines. where high transfection efficiency in a focal region is preferred over systemic delivery. While promising. this approach has traditionally been difficult to administer clinically due to the difficulty in handling and loading of genetic payloads. cytotoxicity of bulk electroporation. and the requirement for investiture in expensive high voltage capital equipment. A more sophisticated approach would incorporate the molecular delivery device and electronics into a pre-loaded disposable patch which can be administered in an out-patient clinical setting. However. this requires scaling the device geometry to enable electroporation and precise dosage dispensation with low voltage electronics. 
 
 The primary objective of the proposed work is the fabrication of an innovative nanopore microneedle (NM) platform for cell transfection through low-power controlled nanoelectroporation (NEP) and electrophoretic insertion of genetic materials. The platform has already been designed. but as cleanroom facilities are required for fabrication. the project has been delayed until funding can be secured. 
 
 The larger study hypothesis of an NSF CAREER proposal submitted by the PI during the summer of 2020 was that an innovative nanopore microneedle (NM) can be used to uniformly disperse genetic material and transfect cells utilizing ultra-low energy electric fields in vitro. However. the NSF review panel stated that. while this research is highly interesting and has great potential for impact. they would like to see the team produce a prototype as preliminary data to de-risk the project. This SRA funding will benefit the re-submission of that application by allowing the fabrication of initial prototypes in fulfillment of this direct request by the review panel. To provide a step beyond the simple meeting of the review panel???s request. the PI has already secured an arrangement with collaborators at North Carolina State University to do characterization experiments (from their own funding) if the UTSA team can produce the initial prototypes.