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Universal intracellular biomolecule delivery with precise dosage control

Cao, Y. ; Chen, H. ; Qiu, R. ; Hanna, M. ; Ma, E. ; Hjort, M. LU orcid ; Zhang, A. ; Lewis, R. S. ; Wu, J. C. LU and Melosh, N. A. (2018) In Science Advances 4(10).
Abstract

Intracellular delivery of mRNA, DNA, and other large macromolecules into cells plays an essential role in an array of biological research and clinical therapies. However, currentmethods yield a wide variation in the amount of material delivered, as well as limitations on the cell types and cargoes possible. Here, we demonstrate quantitatively controlled delivery into a range of primary cells and cell lines with a tight dosage distribution using a nanostrawelectroporation system (NES). In NES, cells are cultured onto track-etched membranes with protruding nanostraws that connect to the fluidic environment beneath the membrane. The tight cell-nanostraw interface focuses applied electric fields to the cell membrane, enabling low-voltage... (More)

Intracellular delivery of mRNA, DNA, and other large macromolecules into cells plays an essential role in an array of biological research and clinical therapies. However, currentmethods yield a wide variation in the amount of material delivered, as well as limitations on the cell types and cargoes possible. Here, we demonstrate quantitatively controlled delivery into a range of primary cells and cell lines with a tight dosage distribution using a nanostrawelectroporation system (NES). In NES, cells are cultured onto track-etched membranes with protruding nanostraws that connect to the fluidic environment beneath the membrane. The tight cell-nanostraw interface focuses applied electric fields to the cell membrane, enabling low-voltage and nondamaging local poration of the cell membrane. Concurrently, the field electrophoretically injects biomolecular cargoes through the nanostraws and into the cell at the same location. We show that the amount of material delivered is precisely controlled by the applied voltage, delivery duration, and reagent concentration. NES is highly effective even for primary cell types or different cell densities, is largely cargo agnostic, and can simultaneously deliver specific ratios of different molecules. Using a simple cell culture well format, the NES delivers into >100,000 cells within 20 s with >95% cell viability, enabling facile, dosage-controlled intracellular delivery for a wide variety of biological applications.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Science Advances
volume
4
issue
10
article number
eaat8131
publisher
American Association for the Advancement of Science (AAAS)
external identifiers
  • scopus:85055861168
  • pmid:30402539
ISSN
2375-2548
DOI
10.1126/sciadv.aat8131
language
English
LU publication?
yes
id
4e16a82a-44d6-4160-b048-9cc8a9ce1328
date added to LUP
2018-11-15 10:01:42
date last changed
2024-04-15 16:34:46
@article{4e16a82a-44d6-4160-b048-9cc8a9ce1328,
  abstract     = {{<p>Intracellular delivery of mRNA, DNA, and other large macromolecules into cells plays an essential role in an array of biological research and clinical therapies. However, currentmethods yield a wide variation in the amount of material delivered, as well as limitations on the cell types and cargoes possible. Here, we demonstrate quantitatively controlled delivery into a range of primary cells and cell lines with a tight dosage distribution using a nanostrawelectroporation system (NES). In NES, cells are cultured onto track-etched membranes with protruding nanostraws that connect to the fluidic environment beneath the membrane. The tight cell-nanostraw interface focuses applied electric fields to the cell membrane, enabling low-voltage and nondamaging local poration of the cell membrane. Concurrently, the field electrophoretically injects biomolecular cargoes through the nanostraws and into the cell at the same location. We show that the amount of material delivered is precisely controlled by the applied voltage, delivery duration, and reagent concentration. NES is highly effective even for primary cell types or different cell densities, is largely cargo agnostic, and can simultaneously deliver specific ratios of different molecules. Using a simple cell culture well format, the NES delivers into &gt;100,000 cells within 20 s with &gt;95% cell viability, enabling facile, dosage-controlled intracellular delivery for a wide variety of biological applications.</p>}},
  author       = {{Cao, Y. and Chen, H. and Qiu, R. and Hanna, M. and Ma, E. and Hjort, M. and Zhang, A. and Lewis, R. S. and Wu, J. C. and Melosh, N. A.}},
  issn         = {{2375-2548}},
  language     = {{eng}},
  number       = {{10}},
  publisher    = {{American Association for the Advancement of Science (AAAS)}},
  series       = {{Science Advances}},
  title        = {{Universal intracellular biomolecule delivery with precise dosage control}},
  url          = {{http://dx.doi.org/10.1126/sciadv.aat8131}},
  doi          = {{10.1126/sciadv.aat8131}},
  volume       = {{4}},
  year         = {{2018}},
}