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Proc Natl Acad Sci U S A. 2019 Apr 16;116(16):7899-7904. doi: 10.1073/pnas.1818553116. Epub 2019 Mar 28.

Nontoxic nanopore electroporation for effective intracellular delivery of biological macromolecules.

Author information

Department of Chemistry, University of California, Berkeley, CA 94720.
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
Department of Materials Science and Engineering, University of California, Berkeley, CA 94720.
Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94705.
Howard Hughes Medical Institute, University of California, Berkeley, CA 94720.
Innovative Genomics Institute, University of California, Berkeley, CA 94720.
Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720.
California Institute for Quantitative Biosciences, Berkeley, CA 94720.
Department of Chemistry, University of California, Berkeley, CA 94720;


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.


electroporation; genome engineering; intracellular delivery; nanopore; nanotechnology


Conflict of interest statement

Conflict of interest statement: Y.C., E.M., J.A.D., and P.Y. are inventors on patent applications (filed by the University of California, Berkeley) related to the nanopore electroporation systems and uses thereof. J.A.D. is a cofounder of Caribou Biosciences, Editas Medicine, Intellia Therapeutics, Scribe Therapeutics, and Mammoth Biosciences and is a member of the board of directors of Driver and Johnson & Johnson and a member of the scientific advisory boards for Caribou Biosciences, Intellia Therapeutics, eFFECTOR Therapeutics, Scribe Therapeutics, Synthego, Metagenomi and Inari. Y.C., S.C.-B., P.Y., and Bozhi Tian are coauthors on a 2018 review article.

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