The Research On LacZ/H2-K～b Gene Transfection With SB Transposon System

PURPOSE The Sleeping Beauty (SB) transposon system canintegrate foreign sequences of DNA in the genome of mouse somatic cellseliciting long-term expression in vivo. This technology holds greatpromise for human gene therapy as a nonviral technology to delivertherapeutic genes. Here, we test the ability of the SB transposon system tofacilitate somatic integration of a transposon containing LacZ with CMVpromoter /H2-K~b with RIP promoter to detect transposase mediatedinsertion of delivered vectors. Our objective is construct a techniqueplatform of transposon system and prepared for larger fragmenttransgenic research with transposon system.METHOD The plasmids, SB transposon and transposase, carryingtarget foreign gene, were packed by DOTAP: CHOL and injected throughhydrodynamaic into adult mice. Then, we analysis the ability to achievelong-term expression in vivo by using SB suggests that somaticintegration of foreign gene using SB is a feasible approach to modelmolecularly defined changes in mouse, and detected it by LacZ staining,histopathology,PCR,RT,Slot Blotting, and the monitoring of thechangge of weight,blood sugar,glucose in urine.RESULT LacZ with CMV promoter carded by SB transposon was found to express in the treated mice. In the mice injected with pancreaticexpressed RIP promoter-H2-K~b gene carried by transposon, there wasdecrease in number and decay in morphology of pancreaticβcell in thehistological analysis of pancreas, decrease of weight, increase of bloodsugar, and other pathological change.CONCLUSION The SB transposon carrying target gene and thetransposase were injected via tail vein into adult mice. And the validity oftransposon system was proved in this study.