Efficient expression of human factor Ⅸ cDNA in liver mediated by hydrodynamics-based plasmid administration

The transfection of plasmid DNA into mammalian cells is an indispensable tool in the study of gene transfer and gene function. Since the original report in 1990 on the successful expression of a reporter gene in muscle[1], plasmid DNA injection has been widely used to mediate gene transfer study. As a gene transfer vector, naked DNA has many obvious advantages. It is easy and cheap to be prepared and more safe after transfection. But the plasmid mediated gene expression is low because of t…     

Efficient expression of human factor Ⅸ cDNA in liver mediated by hydrodynamics-based plasmid administration

Hydrodynamics-based administration via tail vein was used to deliver naked plasmid with human factor IX (hFIX) cDNA in 2.2 mL Ringer‘s solution into mice within 7 s. The peak level of expression of hFIX was 2921 ng/mL in mouse plasma. The hFIX cDNA expression increased with increasing the amount of plasmid DNA injected. The peak level of gene expression declined after repeated injection of plasmid (1459 ng/mL). The hFIX cDNA was detected in various organs, but the highest level of gene expression appeared in liver. Transaminase levels and liver histologicalresults showed that rapid intravenous plasmid injection into mice induced transient focal acute liver damage, which was rapidly repaired within 3--10 d. These results suggested that high-level expression of hFIX cDNA can be achieved by hydrodynamics-based plasmid transfer and this method is nowfurther used for gene therapy and gene function study in our lab.