Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells
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 pl...
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MDPI AG
2016-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/17/3/335 |
| _version_ | 1811283400227553280 |
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| author | Guanglu Liu Yuan Wang Yang Hu Xiaobo Yu Bin Zhu Gaoxue Wang |
| author_facet | Guanglu Liu Yuan Wang Yang Hu Xiaobo Yu Bin Zhu Gaoxue Wang |
| author_sort | Guanglu Liu |
| collection | DOAJ |
| description | 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. |
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| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T02:10:51Z |
| publishDate | 2016-03-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-dad4b74f98504b92a0688d227cd7ad2f2022-12-22T03:07:18ZengMDPI AGInternational Journal of Molecular Sciences1422-00672016-03-0117333510.3390/ijms17030335ijms17030335Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish CellsGuanglu Liu0Yuan Wang1Yang Hu2Xiaobo Yu3Bin Zhu4Gaoxue Wang5College of Science, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaCollege of Animal Science and Technology, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaCollege of Science, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaCollege of Animal Science and Technology, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaCollege of Animal Science and Technology, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaCollege of Animal Science and Technology, Northwest A & F University, Xinong Road 22nd, Yangling 712100, ChinaDNA 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.http://www.mdpi.com/1422-0067/17/3/335DNA vaccinemulti-wall carbon nanotubes (MWCNTs)grass carp reovirus (GCRV)outer capsid protein VP5 |
| spellingShingle | Guanglu Liu Yuan Wang Yang Hu Xiaobo Yu Bin Zhu Gaoxue Wang Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells International Journal of Molecular Sciences DNA vaccine multi-wall carbon nanotubes (MWCNTs) grass carp reovirus (GCRV) outer capsid protein VP5 |
| title | Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells |
| title_full | Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells |
| title_fullStr | Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells |
| title_full_unstemmed | Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells |
| title_short | Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells |
| title_sort | functionalized multi wall carbon nanotubes enhance transfection and expression efficiency of plasmid dna in fish cells |
| topic | DNA vaccine multi-wall carbon nanotubes (MWCNTs) grass carp reovirus (GCRV) outer capsid protein VP5 |
| url | http://www.mdpi.com/1422-0067/17/3/335 |
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