Construction and expression analysis of micro-linear vector as a new general gene therapy vector.
- Author:
Hongsheng WANG
1
;
Xiaoqing LI
;
Yuwen HE
;
Bailu XIE
;
Wenying TANG
;
Jun DU
Author Information
1. Center of Microbiology, Biochemistry and Pharmacology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China.
- Publication Type:Journal Article
- MeSH:
3T3 Cells;
Animals;
CpG Islands;
Genetic Therapy;
methods;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
genetics;
Humans;
Kidney;
cytology;
Mice;
Promoter Regions, Genetic;
genetics;
Transfection;
methods
- From:
Chinese Journal of Biotechnology
2008;24(8):1333-1339
- CountryChina
- Language:Chinese
-
Abstract:
The most difficult field in gene therapy is that vector system should offer both a means of successful transfection and a maximum of safety for the patient. Viral vectors and plasmid vectors are traditional vectors; they may cause unwanted immunological side effects resulting from the expression of nontherapeutic genes. Our aim is to develop a new general gene therapy vector which is suggested to be called as Micro-Linear Vector. The gene expression cassette is capped by our designed cap, including promoter, enhancer, objective gene, and RNA-stabilizing sequence, so it can defend the exnuclease in the eukaryotic cell, at the same time, DNA not encoding the objective gene is reduced to a minimum. The GFP gene is separated from the pEGFP-N3 plasmid, and acts as a reporter gene to construct the Micro-Linear Vector, then both the new vector and the plasmid are transfected to cells, the results are tested by fluorescence microscope and flow cytometry. The results show that the Micro-Linear Vector has a high effective of transfection and safety in 293, 3T3, CNE2 and B95-8 cell lines, at the same time it is less toxicity than the plasmid. We can get the rudiments of conclusion that Micro-Linear Vector has high effection of the transfection and more safety than tradition plasmid in eukaryotic cell.
