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Int. J. Mol. Sci. 2016, 17(3), 335; doi:10.3390/ijms17030335

Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells

1
College of Science, Northwest A & F University, Xinong Road 22nd, Yangling 712100, China
2
College of Animal Science and Technology, Northwest A & F University, Xinong Road 22nd, Yangling 712100, China
*
Author to whom correspondence should be addressed.
Academic Editors: Jun Li and Li Lin
Received: 31 December 2015 / Revised: 29 January 2016 / Accepted: 24 February 2016 / Published: 3 March 2016
(This article belongs to the Special Issue Fish Molecular Biology)
View Full-Text   |   Download PDF [2487 KB, uploaded 3 March 2016]   |  

Abstract

DNA vaccines are considered to be the most promising method against infectious diseases in the aquaculture industry. In the present study, we investigated the potency of ammonium group-functionalized multi-walled carbon nanotubes (MWCNTs) in enhancing the transfection and expression efficiency of plasmid DNA (pEGFP-vp5) in Ctenopharyngodon idellus kidney (CIK) cells. Agarose gel shift assay results show that ammonium group-functionalized carbon nanotubes are able to condense DNA in varying degrees. Scanning electron microscope (SEM) images shows that CIK cells show a great affinity for MWCNTs-NH3+ and the CNTs covering the cell surface tend to orient their tips perpendicularly to the cell surface, and appear to be “needle-pricking the cells”. Transmission electron microscope (TEM) images confirmed that MWCNTs-NH3+ penetrate the cell membranes and are widely dispersed in the CIK cell. Real-time PCR was used to detect the transfection efficiency through the expression of the outer capsid protein (VP5). The results showed that the MWCNTs-NH3+:DNA complexes are able to transfect CIK cells effectively at different charge ratio than naked DNA. Subsequent studies confirmed that both functional groups and charge ratio are important factors that determine the transfection efficiency of plasmid DNA. All these results indicated that MWCNTs-NH3+:DNA complexes could be suitable for developing DNA vaccine for the control of virus infection in the aquaculture industry. View Full-Text
Keywords: DNA vaccine; multi-wall carbon nanotubes (MWCNTs); grass carp reovirus (GCRV); outer capsid protein VP5 DNA vaccine; multi-wall carbon nanotubes (MWCNTs); grass carp reovirus (GCRV); outer capsid protein VP5
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Liu, G.; Wang, Y.; Hu, Y.; Yu, X.; Zhu, B.; Wang, G. Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells. Int. J. Mol. Sci. 2016, 17, 335.

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