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Review
. 2018 Nov 21;23(11):3044.
doi: 10.3390/molecules23113044.

A Review on Electroporation-Based Intracellular Delivery

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Free PMC article
Review

A Review on Electroporation-Based Intracellular Delivery

Junfeng Shi et al. Molecules. .
Free PMC article

Abstract

Intracellular delivery is a critical step in biological discoveries and has been widely utilized in biomedical research. A variety of molecular tools have been developed for cell-based gene therapies, including FDA approved CAR-T immunotherapy, iPSC, cell reprogramming and gene editing. Despite the inspiring results of these applications, intracellular delivery of foreign molecules including nucleic acids and proteins remains challenging. Efficient yet non-invasive delivery of biomolecules in a high-throughput manner has thus long fascinates the scientific community. As one of the most popular non-viral technologies for cell transfection, electroporation has gone through enormous development with the assist of nanotechnology and microfabrication. Emergence of miniatured electroporation system brought up many merits over the weakness of traditional electroporation system, including precise dose control and high cell viability. These new generation of electroporation systems are of considerable importance to expand the biological applications of intracellular delivery, bypassing the potential safety issue of viral vectors. In this review, we will go over the recent progresses in the electroporation-based intracellular delivery and several potential applications of cutting-edge research on the miniatured electroporation, including gene therapy, cellular reprogramming and intracellular probe.

Keywords: electroporation; intracellular delivery; microfabrication; miniatured electroporation.

Conflict of interest statement

The authors state that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Various applications in different fields benefit from the development of molecular tools for intracellular delivery.
Figure 2
Figure 2
General classification of popular intracellular delivery platforms: carrier-mediated delivery strategies (blue background) and popular physical membrane-penetrating delivery methods (green background).
Figure 3
Figure 3
Commonly-used commercial bulk electroporation (BEP) system. From left to right: Gene Pulser Xcell™ from Bio-Rad, Nucleofector™ from Lonza, and Neon® Transfection System from Thermo Fisher Scientific. Figures from Internet. copyright by the companies.
Figure 4
Figure 4
Molecular dynamics showing the progress of an aqueous pore forming within the lipid bilayer during electroporation. From left to right (A) the intact bilayer, (B) a few water molecules enter the lipid regime, starting to form a “water path”, and (C) the neighbouring lipids reorient, stabilizing the “water pore” and allowing the ions to enter. Reprinted with permission from ref. [39] Copyright © 2012, IEEE.
Figure 5
Figure 5
Schematic of electroporation system for cell suspensions.
Figure 6
Figure 6
Representative schematics of micro-scale cargo delivery systems. (A) Electroporation through a micro-fabricated orifice to the measurement of single cell response to external stimuli. Reprinted with permission from ref. [50]. (B) High-throughput micronozzle-based sandwich EP system. Reprinted with permission from ref. [51]. (C) Flow-through comb electroporation device for delivery of macromolecules. Reprinted with permission from ref. [52]. (D) Microfluidic cell squeezing method. Reprinted with permission from ref. [54]. (E) Magnetic tweezers-based 3D MEP for high-throughput gene transfection in living cells. Reprinted with permission from ref. [55].
Figure 7
Figure 7
Representative schematics of nano-scale cargo delivery systems. (A) First-generation Nano-electroporation (NEP) device platform comprised of nanochannel array and optical tweezer for cell loading. Reprinted with permission from ref. [56]. (B) Motivated by limited throughput (i.e., <200 cells) of the proof-of-concecpt NEP system, high-throughput NEP platform, which features a massive nanochannel array in the z-direction handle up to 1 million cells, were developed. Reprinted with permission from ref. [60]. (C) Nanopillar EP. Reprinted with permission from ref. [57]. (D) Nanostraw EP. Reprinted with permission from ref. [58]. (E) Nanofountain probe (NFP) technology based on electroporation and AFM nanotip injection. Reprinted with permission from ref. [59].
Figure 8
Figure 8
Microfabrication of miniaturized electroporation. (A) The fabrication procedure of the silicon 3D NEP chip. (B) Schematic of Bosch Process, deep reactive-ion etching (DRIE) progression. (C) Schematic of different types of optical lithography. Reprinted with permission from ref. [60].
Figure 9
Figure 9
Diagram of gene therapies by personalized immunotherapy. Second row: non-viral transfection of genes into T cells is prerequisite to CAR-T immunotherapy. Reprinted with permission from ref. [65].
Figure 10
Figure 10
TNT-based enhanced reprogramming factor delivery and propagation can rescue whole limbs from necrotizing ischemia. Reprinted with permission from ref. [81].
Figure 11
Figure 11
Monitor intracellular biomarker within individual living cells by molecular beacon probes. (A) micrographs of wild-type Kasumi-1 AML cells transfected with DNMT3A/B MBs. Reprinted with permission from ref. [85]. (B) HeLa cells transfected with MBs and imaged after 24 h of incubation showing that the electroporated cells divided. Reprinted with permission from ref. [86].

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