Date of Award

Winter 2016

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Micro and Nanoscale Systems

First Advisor

Shengnian Wang

Abstract

Electroporation serves as a promising non-viral gene delivery approach, while its current configurations carry drawbacks associated with high-voltage electrical pulses and heterogeneous treatment on individual cells. Here, we developed a new micropillar array electroporation (MAE) platform to advance the delivery of plasmid DNA and RNA to mammalian cells. By introducing well-patterned micropillar array on the electrode surface, the number of pillars each cell faces varies with its cell membrane surface area, despite their large population and random locations. In this way, cell size specific electroporation is conveniently done and contributed to a 2.5~3 fold increase on plasmid DNA transfection and an additional 10-55% knockdown with targeting siRNA, respectively. The delivery efficiency varies with the number and size of the micropillars as well as their pattern density. As MAE works like many single cell electroporation is carried out in a parallel fashion, the electrophysiology response of individual cells is representative, which has the potential to gear up the tedious, cell-specific protocol screening process in the current in vitro bulk electroporation (i.e., electroporation to a large population of cells). Its success might facilitate the wide adoption of electroporation as a safe and effective non-viral gene delivery approach needed in many biological research and clinical treatments.

Download

Share

COinS