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Single molecule delivery into living cells

Author

Listed:
  • Chalmers C. Chau

    (University of Leeds
    University of Leeds
    University of Leeds)

  • Christopher M. Maffeo

    (University of Illinois at Urbana–Champaign
    University of Illinois at Urbana-Champaign)

  • Aleksei Aksimentiev

    (University of Illinois at Urbana–Champaign
    University of Illinois at Urbana-Champaign)

  • Sheena E. Radford

    (University of Leeds)

  • Eric W. Hewitt

    (University of Leeds)

  • Paolo Actis

    (University of Leeds
    University of Leeds)

Abstract

Controlled manipulation of cultured cells by delivery of exogenous macromolecules is a cornerstone of experimental biology. Here we describe a platform that uses nanopipettes to deliver defined numbers of macromolecules into cultured cell lines and primary cells at single molecule resolution. In the nanoinjection platform, the nanopipette is used as both a scanning ion conductance microscope (SICM) probe and an injection probe. The SICM is used to position the nanopipette above the cell surface before the nanopipette is inserted into the cell into a defined location and to a predefined depth. We demonstrate that the nanoinjection platform enables the quantitative delivery of DNA, globular proteins, and protein fibrils into cells with single molecule resolution and that delivery results in a phenotypic change in the cell that depends on the identity of the molecules introduced. Using experiments and computational modeling, we also show that macromolecular crowding in the cell increases the signal-to-noise ratio for the detection of translocation events, thus the cell itself enhances the detection of the molecules delivered.

Suggested Citation

  • Chalmers C. Chau & Christopher M. Maffeo & Aleksei Aksimentiev & Sheena E. Radford & Eric W. Hewitt & Paolo Actis, 2024. "Single molecule delivery into living cells," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-48608-3
    DOI: 10.1038/s41467-024-48608-3
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    References listed on IDEAS

    as
    1. Mukhil Raveendran & Andrew J. Lee & Rajan Sharma & Christoph Wälti & Paolo Actis, 2020. "Rational design of DNA nanostructures for single molecule biosensing," Nature Communications, Nature, vol. 11(1), pages 1-9, December.
    2. Roberta Cascella & Serene W. Chen & Alessandra Bigi & José D. Camino & Catherine K. Xu & Christopher M. Dobson & Fabrizio Chiti & Nunilo Cremades & Cristina Cecchi, 2021. "The release of toxic oligomers from α-synuclein fibrils induces dysfunction in neuronal cells," Nature Communications, Nature, vol. 12(1), pages 1-16, December.
    3. Sjors H. W. Scheres & Benjamin Ryskeldi-Falcon & Michel Goedert, 2023. "Molecular pathology of neurodegenerative diseases by cryo-EM of amyloids," Nature, Nature, vol. 621(7980), pages 701-710, September.
    4. Duverney Chaverra-Rodriguez & Vanessa M. Macias & Grant L. Hughes & Sujit Pujhari & Yasutsugu Suzuki & David R. Peterson & Donghun Kim & Sage McKeand & Jason L. Rasgon, 2018. "Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing," Nature Communications, Nature, vol. 9(1), pages 1-11, December.
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