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Related Experiment Videos

Vector-free intracellular delivery by reversible permeabilization.

Shirley O'Dea1, Valeria Annibaldi1, Louise Gallagher1

  • 1Avectas Ltd., Maynooth, Co. Kildare, Ireland.

Plos One
|March 31, 2017
PubMed
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A novel vector-free method uses reversible cell permeabilization with ethanol to rapidly deliver diverse cargos like proteins and mRNA into various cell types. This gentle, efficient technique maintains cell viability and shows potential for broad research and clinical applications.

Area of Science:

  • Cell biology
  • Biotechnology
  • Drug delivery

Background:

  • Efficient intracellular delivery is crucial for biological research and therapeutics.
  • Existing vector-free methods often face limitations in cargo versatility, cell type applicability, or cell viability.
  • There is a need for robust, non-viral intracellular delivery systems.

Purpose of the Study:

  • To develop and validate a novel, vector-free method for rapid intracellular delivery of diverse cargos.
  • To demonstrate the efficiency, versatility, and low toxicity of this new delivery approach.
  • To establish a method compatible with various cell types and high-throughput applications.

Main Methods:

  • Developed a vector-free delivery solution using low-level ethanol for reversible cell permeabilization.

Related Experiment Videos

  • Utilized controlled temporal and volumetric contact with the delivery solution for cargo introduction.
  • Employed an atomizer for direct application of cargo and delivery solution to cells in multi-well plates.
  • Reversed permeabilization using phosphate-buffered saline followed by standard cell culture medium.

    • Main Results:

    • Successfully delivered proteins, mRNA, and plasmid DNA into various cell types, including primary and difficult-to-transfect cells.
    • Confirmed cargo functionality post-delivery in proof-of-principle studies.
    • Demonstrated co-delivery of different cargo types with low observed toxicity.
    • Achieved endocytic-independent, cytoplasmic delivery in situ for both adherent and suspension cells.

    Conclusions:

    • The developed method offers a rapid, gentle, and highly reproducible means of intracellular cargo delivery.
    • This vector-free approach is versatile, accommodating various cargo types and cell lines while maintaining cell viability.
    • The technique's compatibility with high-throughput assays and potential for drug discovery and clinical applications is significant.