[Construction and expression analysis of micro-linear vector as a new general gene therapy vector]

Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1333-9.
[Article in 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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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*

Substances

  • Green Fluorescent Proteins