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Abstract A high‐throughput non‐viral intracellular delivery platform is introduced for the transfection of large cargos with dosage‐control. This platform, termed Acoustic‐Electric Shear Orbiting Poration (AESOP), optimizes the delivery of intended cargo sizes with poration of the cell membranes via mechanical shear followed by the modulated expansion of these nanopores via electric field. Furthermore, AESOP utilizes acoustic microstreaming vortices wherein up to millions of cells are trapped and mixed uniformly with exogenous cargos, enabling the delivery of cargos into cells with targeted dosages. Intracellular delivery of a wide range of molecule sizes (<1 kDa to 2 MDa) with high efficiency (>90%), cell viability (>80%), and uniform dosages (<60% coefficient of variation (CV)) simultaneously into 1 million cells min−1per single chip is demonstrated. AESOP is successfully applied to two gene editing applications that require the delivery of large plasmids: i) enhanced green fluorescent protein (eGFP) plasmid (6.1 kbp) transfection, and ii) clustered regularly interspaced short palindromic repeats (CRISPR)‐Cas9‐mediated gene knockout using a 9.3 kbp plasmid DNA encoding Cas9 protein and single guide RNA (sgRNA). Compared to alternative platforms, this platform offers dosage‐controlled intracellular delivery of large plasmids simultaneously to large populations of cells while maintaining cell viability at comparable delivery efficiencies.
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Laser-Activated Self-Assembled Thermoplasmonic Nanocavity Substrates for Intracellular Delivery
Intracellular delivery is crucial for cellular engineering and the development of therapeutics. Laser-activated thermoplasmonic nanostructured surfaces are a promising platform for high-efficiency, high-viability, high-throughput intracellular delivery. Their fabrication, however, typically involves complicated nanofabrication techniques, limiting the approach’s applicability. Here, colloidal self-assembly and templating are used to fabricate large arrays of thermoplasmonic nanocavities simply and cost-effectively. These laser-activated substrates are used to deliver membrane-impermeable dye into cells at an efficiency of 78% and throughput of 30 000 cells min–1 while maintaining 87% cell viability. Proof-of-concept data show delivery of large cargoes ranging from 0.6 to 2000 kDa to cells without compromising viability.
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- Award ID(s):
- 1806434
- PAR ID:
- 10560751
- Publisher / Repository:
- ACS
- Date Published:
- Journal Name:
- ACS Applied Bio Materials
- Volume:
- 1
- Issue:
- 6
- ISSN:
- 2576-6422
- Page Range / eLocation ID:
- 1793 to 1799
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1021/acsabm.8b00447
