[The transfer and expression of human clotting factor IX in muscle mediated by electroporation].
H Wang, F Wang, L Chen, J Xue, Dan Lu
Abstract
H Wang, F Wang, L Chen, J Xue, Dan Lu
Abstract
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.
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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.
Key concepts: Electroporation, Molecular biology, Complementary DNA, Plasmid, Transfection, Recombinant DNA, Skeletal muscle, Biology