Abstract
Electroporation serves as a promising non-viral gene delivery approach, while its current configuration carries several 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 electroporation-based delivery of DNA and RNA probes into mammalian cells. By introducing well-patterned micropillar array texture on the electrode surface, the number of pillars each cell faces varies with its plasma membrane surface area, despite their large population and random locations. In this way, cell size specific electroporation is conveniently carried out, contributing to a 2.5~3 fold increase on plasmid DNA transfection and an additional 10–55% transgene knockdown with siRNA probes, respectively. The delivery efficiency varies with the number and size of micropillars as well as their pattern density. As MAE works like many single cell electroporation are carried out in parallel, the electrophysiology response of individual cells is representative, which has potentials to facilitate the tedious, cell-specific protocol screening process in current bulk electroporation (i.e., electroporation to a large population of cells). Its success might promote the wide adoption of electroporation as a safe and effective non-viral gene delivery approach needed in many biological research and clinical treatments.
Highlights
Gene induction and/or inhibition provide powerful tools to understand gene functions[1], control cellular signals[2], and develop new therapeutic technologies[3]
Successful transfection is observed in all three cases with many cells expressed green fluorescence protein (GFP) (Fig. 2a and Supp Figure S1)
These results confirm the enhancement of Micropillar Array Electroporation (MAE) on plasmid transfection to mammalian cells and the improvement is attributed to both the micropillar features and the closely placed electrode configuration
Summary
Array Electroporation received: 09 March 2015 accepted: 14 November 2016 Published: 07 December 2016. Unlike some pioneer work in which a few micro- or nanoscale pillar electrodes were used as the replacement of capillary electrodes to monitor the intracellular electrical signals of single or a few cells for electrophysiology study (i.e., SCE)[54,55], this new MAE setup utilizes well-patterned, large-scale (centimeter size) micropillar array to achieve size specific treatment to cells of a large population for efficient uptake of exogenous payload (like BE). It works like many SCE units are carried out in parallel with no need for cell positioning. Several adherent and suspension cell lines including some hard-to-transfect cells and stem cells were tested to demonstrate its broad effectiveness
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