Nanochannel electroporation delivers precise amounts of biomolecules into living cells

Abstract

Many transfection techniques can deliver biomolecules into cells, but the dose cannot be controlled precisely. Delivering well-defined amounts of materials into cells is important for various biological studies and therapeutic applications. Here, we show that nanochannel electroporation can deliver precise amounts of a variety of transfection agents into living cells. The device consists of two microchannels connected by a nanochannel. The cell to be transfected is positioned in one microchannel using optical tweezers, and the transfection agent is located in the second microchannel. Delivering a voltage pulse between the microchannels produces an intense electric field over a very small area on the cell membrane, allowing a precise amount of transfection agent to be electrophoretically driven through the nanochannel, the cell membrane and into the cell cytoplasm, without affecting cell viability. Dose control is achieved by adjusting the duration and number of pulses. The nanochannel electroporation device is expected to have high-throughput delivery applications.

In vitro and ex vivo strategies for intracellular delivery

“…This innovation enables the generation of a single large hole rather than the numerous small pores characteristic of conventional electroporation, which is less amenable to free passage of large materials. For example, conventional electroporation appears to exploit the charge of nucleic acids, such as plasmids and mRNA, in order to partially embed them in pores, resulting in subsequent internalization through active membrane-trafficking pathways rather than direct delivery 81 . In contrast, nanochannel electroporation achieves improved dose control, enhanced electrophoretic delivery deeper into the cell, and the ability to deliver materials that bulk electroporation often struggles to deliver, such as quantum dots.…”

miRNA Control of Tissue Repair and Regeneration

“…With the use of an optical tweezer, the target cell is positioned in one microchannel, and the miRNA of interest may be placed in the second microchannel. 95 The production of an intense electric field is guided by the introduction of a voltage pulse between microchannels directed at a very small area on the cell membrane. This enables controlled delivery of specific amounts of oligonucleotide driven electrophoretically through the nanochannel, cell membrane, and into the cytoplasm while defending cell viability.…”

“…This enables controlled delivery of specific amounts of oligonucleotide driven electrophoretically through the nanochannel, cell membrane, and into the cytoplasm while defending cell viability. 95 Variations in the duration of nanoelectroporation and number of pulses applied may help control dosage of cargo delivery. Although there is no reported in vivo evidence miRNA and Regenerative Medicine…”

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

“…To use this dimensional advantage, an electric field can easily intense in the local region of the single cell membrane, by which high transfection rate and high cell viability were achieved [27,29]. However localized electroporation can also be performed by nanochannel ion transportation using the electrophoresis method [30]. To apply electrical field in local region of the single cell membrane, the process is known as localized single cell membrane electroporation (LSCMEP), by which precise and controllable drug delivery is possible [27,29,30].…”