| 摘要 |
Highly efficient regulated promoters (RPs) and transactivators are provided. The RPs contain 2-8 transactivator binding domains positioned in an optimized manner with respect to each other. In a tetracycline-regulatable system, the RP displays 5- to 10-fold enhanced regulation efficiency by maintaining high maximal expression while offering 5- to 10-fold reduced basal leakiness. In transient studies, these novel promoters display over 900-fold gene regulation at 1:1 ratio of transactivator to reporter plasmid. Furthermore, these promoters preserve their regulator efficienty in the context of a single positive feedback regulatory vector that presents ease of delivery of the system for many uses, including for use in vivo and in gene therapy. Finally, humanized transactivators are provided, for example a tetracycline transactivator utilizing the human transactivational domain of NF- kappa p65 protein fused to tetracycline repressor (terR) is provided that functions as efficiently as the tetracycline-regulated transactivator (tTA) and should reduce the potential immunogenicity of the original tTA. |
