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I.-J. Wang, M.-C. Jhuang, Y.-H. Chen, L.-K. Yeh, T.-H. Yang; Transfect Bovine Cornea Epithelial Cells With Chitosan-Modified Adenovirus. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2729.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the possibility to increase the transfection efficiency with chitosan-linked adenovirus in bovine cornea epithelial cells.
Adenovirus(Ad) was modified with a heterogeneous crosslinker(GMBS), and the amino group of chitosan was modified with 2-iminothiolane. Then the chitosan-SH was reacted with GMBS-Ad via thioether. The size and zeta-potential of chitosan-modified Ad were measured. HEK293 and bovine cornea epithelial cells were transfected with adenovirus and chitosan-modified Ad.
Analysis of particle size showed an average diameter of the unmodified Ad of 120.0nm, compared with 139.5nm for the chitosan-modified Ad. Zeta-potential showed -24.9±6.4 mV for the unmodified Ad, compared with 2.5±3.7 mV for the chitosan-modified Ad. When infected, HEK293 and bovine cornea epithelial cells with the unmodified Ad appeared bright fluorescence in over 90% of cells at 24 h after infection, while the chitosan-modified Ad gave no detectable fluorescence signal.
The ability of adenovirus to infect cells was essentially ablated by modification of adenovirus with chitosan.
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