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Nontoxic nanopore electroporation for effective intracellular delivery of biological macromolecules.

ABSTRACT: 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.

SUBMITTER: Cao Y 

PROVIDER: S-EPMC6475394 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Nontoxic nanopore electroporation for effective intracellular delivery of biological macromolecules.

Cao Yuhong Y   Cao Yuhong Y   Ma Enbo E   Cestellos-Blanco Stefano S   Zhang Bei B   Qiu Ruoyi R   Su Yude Y   Doudna Jennifer A JA   Yang Peidong P  

Proceedings of the National Academy of Sciences of the United States of America 20190328 16

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-p  ...[more]

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