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Acoustically-mediated intracellular delivery.

Shwathy Ramesan1, Amgad R Rezk1, Chaitali Dekiwadia2

  • 1Micro/Nanophysics Research Laboratory, School of Engineering, RMIT University, Melbourne, VIC 3000, Australia. leslie.yeo@rmit.edu.au.

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Summary
This summary is machine-generated.

High-frequency ultrasound rapidly enhances gene delivery into cells by temporarily reorganizing cell membranes. This novel acoustic excitation method improves therapeutic molecule uptake while maintaining high cell viability (>97%) and efficient gene silencing.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Biophysics

Background:

  • Efficient ex vivo cell engineering is crucial for gene editing therapies.
  • Current intracellular delivery methods face challenges with cell viability and endosomal escape.

Purpose of the Study:

  • To develop a rapid and efficient method for intracellular delivery of therapeutic molecules into patient-harvested cells.
  • To improve cell viability and cargo distribution compared to existing techniques.

Main Methods:

  • Exposure of cells to high-frequency (>10 MHz) acoustic excitation for a short duration (10 minutes).
  • Utilizing gold nanoparticles and therapeutic molecules for delivery.
  • Assessing cell viability and cargo distribution via microscopy and functional assays.

Main Results:

  • Acoustic excitation temporarily reorganizes the cell membrane, enhancing molecule translocation.
  • High cell viability (>97%) and rapid cell self-healing were observed.
  • Internalized cargo was uniformly distributed in the cytosol, bypassing endosomal escape issues.
  • siRNA delivery resulted in over twofold enhancement in gene silencing.

Conclusions:

  • High-frequency acoustic excitation offers a promising, non-invasive method for efficient ex vivo cell engineering.
  • This technique significantly improves intracellular delivery of therapeutics, enhancing gene silencing efficacy.
  • The method's high cell viability and efficient cargo delivery demonstrate its potential for clinical applications in gene therapy.