Electric Shock Mediated By Sleeping Beauty-transposon-mediated Human Coagulation Factor Ix Gene Expression

(1)Gene transfer and expression mediated by electroporation via mouse muscleTo probe into the feasibility of increasing hFIX cDNA transfer and expression in muscle, both Lac-Z-expressiong plasmid pCMVβ and hFIX-expressing plasmid pCMV-hFIX were transferd into mouse muscle by in vivo electroporation in low-frequency and moderate electric fields to prmote their transfer and expression. The highest expression level of hFIX in mice with electroporation was 24.4±3.9ng/mL, more than 4 times higher than that of mice without electroporation(5.6±2.3ng/mL). It was proved that electric stimulation could significantly increase the transfer and expression of hFIX in mouse muscle.(2)Study of hFIX expression mediated by Sleeping Beauty transposonSleeping Beauty transposon vectors expressing hFIX (pSB-hFIX) and transposase expressing plasmids (pCMV-SB10) were both injected into mice by hydrodynamics-based administration via tail vein. Effects of different ratio and concentration of pSB-hFIX and the pCMV-SB10 were investigated to optimize hFIX expression. The expression of hFIX in mouse plasma lasted for more than 4 month, and the highest expression level of hFIX reached 6475.6±278.1ng/mL. It was demonstrated that long-term and high expression of hFIX in mice could be achieved by using SB transposon system.