Research On Preparation Of Silica Based Vectors And Transfection In Vivo

This paper was aimed to prepare organic-inorganic hybrid nano-gene vector based on silica nano materials and cation polymers for gene delivery.The following three aspects were discussed in this paper:1.CMG nanoparticles were synthesized through a rapid one-step aqueous synthetic approach with carboxymethylchitosan,GPSM and ammonia system.The morphology,size and surface potential were characterized by TEM and Nano-ZS Zetasizer.The structure was determined by FTIR and TGA.KALA peptide was modified to the surface of CMG nanoparticles to enhance its transfection efficiency as a non-viral vector.Through the analysis of FTIR,CLSM and fluorescence spectrometry,KALA was demonstrated to connected CMG nanoparticles successfully.Agarose gel electrophoresis showed that the CMG-KALA nanoparticles can protect DNA from degration.Thus,CMG-KALA nanoparticles have great potential as gene vectors for transfection in vitro.2.GS-peptide nanoparticles were synthesized as previous reported method in order to improve the ability of cell penetrating,endosome escaping and nuclear targeting.The connection ratio was detected by fluorescence spectra.Size and surface potential was detected by Nano-ZS Zetasizer.Through CLSM and agarose gel electrophoresis DNA bonding ability of GS-peptide was characterized.Based on the prediction of secondary structure of peptide with DNAstar biological activity and mechanism of connection was discussed.3.Based on previous studies of GS-Tat transfection in vitro,further study was carried out in vivo.Biodistribution and gene express were detected respectively. The results will demonstrate the feasibility of GS-Tat nanoparticles used as a gene deliver system.