title: Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells.
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.
authors.affiliation: College of Science, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. 563310592@163.com.
authors.forename: Guanglu
authors.initials: G
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. 18792426961@163.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Science, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. 18729316235@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hu
authors.affiliation: College of Animal Science and Technology, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. yuxiaobo6@126.com.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Yu
authors.affiliation: College of Animal Science and Technology, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. zhubin1227@126.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhu
authors.affiliation: College of Animal Science and Technology, Northwest A &amp; F University, Xinong Road 22nd, Yangling 712100, China. wanggaoxue@126.com.
authors.forename: Gaoxue
authors.initials: G
authors.lastname: Wang
mesh_terms: D064751:Ammonium Compounds / Q000737:chemistry*; D000818:Animals; D017756:Aquaculture; D036022:Capsid Proteins / Q000235:genetics; D002347:Carps / Q000235:genetics* / Q000821:virology; D002478:Cells, Cultured; D005393:Fish Diseases / Q000517:prevention & control / Q000821:virology; D015870:Gene Expression; D007668:Kidney / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D037742:Nanotubes, Carbon / Q000737:chemistry*; D030781:Organisms, Genetically Modified; D010957:Plasmids / Q000235:genetics* / Q000648:ultrastructure; D014162:Transfection; D019444:Vaccines, DNA / Q000235:genetics
chemical_list: D064751:Ammonium Compounds; D036022:Capsid Proteins; D037742:Nanotubes, Carbon; D019444:Vaccines, DNA
keywords: DNA vaccine; grass carp reovirus (GCRV); multi-wall carbon nanotubes (MWCNTs); outer capsid protein VP5