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Nontoxic nanopore electroporation for effective intracellular delivery of biological macromolecules.
Cao, Yuhong; Ma, Enbo; Cestellos-Blanco, Stefano; Zhang, Bei; Qiu, Ruoyi; Su, Yude; Doudna, Jennifer A; Yang, Peidong.
Affiliation
  • Cao Y; Department of Chemistry, University of California, Berkeley, CA 94720.
  • Ma E; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
  • Cestellos-Blanco S; Department of Materials Science and Engineering, University of California, Berkeley, CA 94720.
  • Zhang B; Department of Chemistry, University of California, Berkeley, CA 94720.
  • Qiu R; Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94705.
  • Su Y; Department of Chemistry, University of California, Berkeley, CA 94720.
  • Doudna JA; Department of Chemistry, University of California, Berkeley, CA 94720.
  • Yang P; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
Proc Natl Acad Sci U S A ; 116(16): 7899-7904, 2019 04 16.
Article in En | MEDLINE | ID: mdl-30923112
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Transfection / Cytological Techniques / Electroporation / Nanopores / Gene Editing Limits: Humans Language: En Journal: Proc Natl Acad Sci U S A Year: 2019 Document type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Transfection / Cytological Techniques / Electroporation / Nanopores / Gene Editing Limits: Humans Language: En Journal: Proc Natl Acad Sci U S A Year: 2019 Document type: Article