Telomerase is a particular reverse transcriptase that not only synthesizes and maintains the telomere but also promotes the proliferation of resting cells and prevents cellular senescence. The advantages of the Sleeping Beauty transposon system include prolonged transgene expression without eliciting an immunogenic response, no possibility of RCV and ease of construction. Tissue-specific therapeutic gene expression is extremely important in gene therapy, because non-specific expression can cause an immune response of the transduced cells that can severely limit the stability of the transgene. The SB system containing the telomerase gene controlled by two chimeric transthyretin (TTR) gene promoters/enhancers, the human alcohol dehydrogenase gene promoter (ADHp), and the SV40 viral enhancer (SV40VE) was constructed in order to activate hepatocyte cell growth. The higher expression was achieved using these elements and FACS analysis showed that this system was effective in hepatocyte targeted gene therapy. Our new SB mediated telomerase delivery system for hepatocytes can be used in human gene therapy applications.