Abstract
Cyclic stretch has been shown to alter cell physiology, cytoskeletal structure, signal transduction, and gene expression in a variety of cell types. To determine the effects of stretch on the gene transfer process, we compared the transfection efficiencies of human A549 cells grown either statically or exposed to 10% cyclic stretch (Δ surface area) at 60 cycles/min (1 Hz) for 24 hours prior to, and/or after transfection with pEGFP-N1 and pCMV-lux-DTS using lipoplex or electroporation. Stretching the cells prior to transfection had no effect on gene transfer. By contrast, cyclic, but not continuous, stretch applied immediately after transfection for as little as 30 minutes resulted in a 10-fold increase in gene transfer and expression by either transfection technique. These stretch conditions did not result in rupture of the plasma membrane based on the fact that DNA was unable to enter stretched cells unless either an electric field was applied or the DNA was complexed with liposomes. Taken together with the timing of the stretch response and the known effects of stretch on transcription, these findings suggest that cyclic stretch may be altering the intracellular transport of plasmids to increase gene expression.
Original language | English (US) |
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Pages (from-to) | 542-549 |
Number of pages | 8 |
Journal | Molecular Therapy |
Volume | 7 |
Issue number | 4 |
DOIs | |
State | Published - Apr 1 2003 |
Funding
We would like to thank Jacob I. Sznajder and Jennifer L. Young for stimulating discussions. This work was supported in part by a grant from the Institute for Bioengineering and Nanoscience in Advanced Medicine of Northwestern University, the Crane Asthma Foundation, and NIH grants HL59956 and EY12962 (DAD).
Keywords
- Cyclic stretch
- Electroporation
- Gene delivery
- Gene expression
- Gene therapy
- Lipoplex
- Nonviral vectors
- Plasmid DNA
- Transfection
ASJC Scopus subject areas
- Molecular Medicine
- Molecular Biology
- Genetics
- Pharmacology
- Drug Discovery