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[The transfer and expression of human clotting factor IX in muscle mediated by electroporation]
1School of Sciences, Institute of Genetics, Fudan University, Shanghai, 200433 P.R.China. darlu@fudan.edu.cn
Objective:
To probe into the feasibility of increasing hFIX cDNA transfer and expression in muscle.
Methods:
The high-frequency electric field was used to promote both Lac-Z-encoding plasmid pCMV beta and hFIX-expressing plasmid G1NaMCIX to transfer and express in muscle. The effects of frequency and length of square pulse, as well as eletroporation time on hFIX expression were investigated.
Results:
Electric stimulation could increase the transfer and expression of pCMV beta in muscle, the number of X-gal positive myofiber cells in electroporation-treated mice is 2.1 times larger than that of mice not treated by electroporation (P<0.01). The most optimal electric simulation condition for hFIX cDNA transfer and expression was obtained, under this condition, the highest level of hFIX antigen in plasma is (40+/- 5.4) ng/ml and 7 times higher than that of mice without electroporation P < 0.001).
Conclusion:
Electroporation is able to enhance hFIX cDNA transfer and expression in muscle efficiently.