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Quantitative Analysis of Plasmid DNA Distribution and Transgene Expression Following Cell Squeezing.

Justin Sylvers¹, Joseph Krakowiecki¹, Yutong Wu¹

¹Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA.

Annals of Biomedical Engineering

|July 11, 2026

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Microfluidic cell squeezing enhances DNA delivery but reduces gene expression due to chromatin condensation. Inhibiting chromatin condensation may improve gene delivery efficiency.

Area of Science:

Biotechnology
Cell Biology
Molecular Medicine

Background:

Microfluidic cell squeezing is a mechanoporation technique for macromolecule delivery.
Mechanoporation alone has limitations for efficient gene delivery.
Integrating electrotransfection with mechanoporation offers a potential gene delivery strategy.

Purpose of the Study:

To develop and evaluate a gene delivery strategy combining electrotransfection and microfluidic cell squeezing.
To investigate the impact of mechanoporation on intracellular DNA transport and transgene expression.

Main Methods:

Fabrication of a microfluidic device for controlled cell squeezing.
Electrotransfection of mammalian cells with plasmid DNA (pDNA).
Assessment of intracellular pDNA transport and transgene expression in squeezed versus un-squeezed cells.

Keywords:

Cell squeezing DNA nuclear entry Endocytosis Mechanoporation Microfluidics Nuclear deformation

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Main Results:

Cell squeezing increased pDNA cellular uptake and nuclear accumulation by over eightfold.
Despite enhanced DNA delivery, transgene mRNA levels were unchanged.
Protein production was reduced in squeezed cells, potentially due to squeezing-induced chromatin condensation.

Conclusions:

Chromatin condensation induced by cell squeezing limits transgene expression.
Pharmacological inhibition of chromatin condensation could enhance mechanoporation-facilitated gene delivery.