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Chinese Journal of Biotechnology ; (12): 1333-1339, 2008.
Artículo en Chino | WPRIM | ID: wpr-275381

RESUMEN

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.


Asunto(s)
Animales , Humanos , Ratones , Células 3T3 , Islas de CpG , Terapia Genética , Métodos , Vectores Genéticos , Genética , Proteínas Fluorescentes Verdes , Genética , Riñón , Biología Celular , Regiones Promotoras Genéticas , Genética , Transfección , Métodos
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