Alexander Helen K, Booy Evan P, Xiao Wenyan, Ezzati Peyman, Baust Heinrich, Los Marek
Cancer Care Manitoba, Manitoba Institute of Cell Biology, Winnipeg, Canada.
Arch Immunol Ther Exp (Warsz). 2007 May-Jun;55(3):139-49. doi: 10.1007/s00005-007-0025-7. Epub 2007 Jun 8.
"On-demand" regulation of gene expression is a powerful tool to elucidate the functions of proteins and biologically-active RNAs. We describe here three different approaches to the regulation of expression or activity of genes or proteins. Promoter-based regulation of gene expression was among the most rapidly developing techniques in the 1980s and 1990 s. Here we provide basic information and also some characteristics of the metallothionein-promoter-based system, the tet-off system, Muristerone-A-regulated expression through the ecdysone response element, RheoSwitch, coumermycin/novobiocin-regulated gene expression, chemical dimerizer-based promoter activation systems, the "Dual Drug Control" system, "constitutive androstane receptor"-based regulation of gene expression, and RU486/mifepristone-driven regulation of promoter activity. A large part of the review concentrates on the principles and usage of various RNA interference techniques (RNAi: siRNA, shRNA, and miRNA-based methods). Finally, the last part of the review deals with historically the oldest, but still widely used, methods of temperature-dependent regulation of enzymatic activity or protein stability (temperature-sensitive mutants). Due to space limitations we do not describe in detail but just mention the tet-regulated systems and also fusion-protein-based regulation of protein activity, such as estrogen-receptor fusion proteins. The information provided below is aimed to assist researchers in choosing the most appropriate method for the planned development of experimental systems with regulated expression or activity of studied proteins.
基因表达的“按需”调控是阐明蛋白质和生物活性RNA功能的强大工具。我们在此描述三种调控基因或蛋白质表达或活性的不同方法。基于启动子的基因表达调控是20世纪80年代和90年代发展最为迅速的技术之一。在此,我们提供基于金属硫蛋白启动子的系统、四环素调控关闭系统、通过蜕皮激素反应元件由米非司酮调控的表达、RheoSwitch、香豆霉素/新生霉素调控的基因表达、基于化学二聚体的启动子激活系统、“双药控制”系统、基于“组成型雄甾烷受体”的基因表达调控以及RU486/米非司酮驱动的启动子活性调控的基本信息和一些特点。综述的很大一部分集中在各种RNA干扰技术(RNAi:基于小干扰RNA、短发夹RNA和微小RNA的方法)的原理和应用上。最后,综述的最后一部分涉及历史上最古老但仍广泛使用的温度依赖性调控酶活性或蛋白质稳定性的方法(温度敏感突变体)。由于篇幅限制,我们不详细描述但仅提及四环素调控系统以及基于融合蛋白的蛋白质活性调控,如雌激素受体融合蛋白。以下提供的信息旨在帮助研究人员为计划开发的具有所研究蛋白质调控表达或活性的实验系统选择最合适的方法。