Journal Article

Nontoxic nanopore electroporation for effective intracellular delivery of biological macromolecules

Qiu, Ruoyi, Cao, Yuhong, Cestellos-Blanco, Stefano, Yang, Peidong, Zhang, Bei, Ma, Enbo, Su, Yude, Doudna, Jennifer A.

2019

Overview

We present a simple nanopore-electroporation (NanoEP) platform for delivery of nucleic acids, functional protein, and Cas9 single-guide RNA ribonucleoproteins into both adherent and suspension cells with up to 80% delivery efficiency and >95% cell viability. Low-voltage electric pulses permeabilize a small area of cell membrane as a cell comes into close contact with the nanopores. The biomolecule cargo is then electrophoretically drawn into the cells through the nanopores. In addition to high-performance delivery with low cell toxicity, the NanoEP system does not require specialized buffers, expensive materials, complicated fabrication processes, or cell manipulation; it simply consists of a generic nanopore-embedded water-filter membrane and a low-voltage square-wave generator. Ultimately, the NanoEP platform offers an effective and flexible method for universal intracellular delivery.

• Publisher: National Academy of Sciences
• Subject: BASIC BIOLOGICAL SCIENCES / Biological Sciences / Biomolecules / Cell Biology / Cell membranes / Cell Survival / Cell viability / Cytological Techniques - methods / Electric contacts / Electric potential / Electric pulses / Electroporation / Electroporation - instrumentation / Electroporation - methods / Engineering / Equipment Design / Fabrication / Gene Editing - methods / genome engineering / HeLa Cells / Humans / Intracellular / intracellular delivery / Intracellular Space - metabolism / Macromolecules / nanopore / Nanopores / nanotechnology / Nucleic acids / Physical Sciences / Porosity / Recombinant Proteins - genetics / Recombinant Proteins - metabolism / Ribonucleic acid / Ribonucleoproteins / RNA / Toxicity / Transfection - methods / Voltage / Water purification