Christophorou Maria A, Martin-Zanca Dionisio, Soucek Laura, Lawlor Elizabeth R, Brown-Swigart Lamorna, Verschuren Emmy W, Evan Gerard I
Cancer Research Institute, University of California San Francisco Comprehensive Cancer Center, San Francisco, California 94143-0875, USA.
Nat Genet. 2005 Jul;37(7):718-26. doi: 10.1038/ng1572. Epub 2005 May 29.
To investigate the functions of the p53 tumor suppressor, we created a new knock-in gene replacement mouse model in which the endogenous Trp53 gene is substituted by one encoding p53ER(TAM), a p53 fusion protein whose function is completely dependent on ectopic provision of 4-hydroxytamoxifen. We show here that both tissues in vivo and cells in vitro derived from such mice can be rapidly toggled between wild-type and p53 knockout states. Using this rapid perturbation model, we define the kinetics, dependence, persistence and reversibility of p53-mediated responses to DNA damage in tissues in vivo and to activation of the Ras oncoprotein and stress in vitro. This is the first example to our knowledge of a new class of genetic model that allows the specific, rapid and reversible perturbation of the function of a single endogenous gene in vivo.
为了研究p53肿瘤抑制因子的功能，我们创建了一种新的基因敲入替换小鼠模型，其中内源性Trp53基因被一个编码p53ER(TAM)的基因所取代，p53ER(TAM)是一种p53融合蛋白，其功能完全依赖于异位提供4-羟基他莫昔芬。我们在此表明，源自此类小鼠的体内组织和体外细胞都可以在野生型和p53基因敲除状态之间快速切换。利用这个快速扰动模型，我们定义了p53介导的对体内组织DNA损伤以及体外Ras癌蛋白激活和应激反应的动力学、依赖性、持续性和可逆性。据我们所知，这是新型遗传模型的首个实例，该模型能够在体内对单个内源性基因的功能进行特异性、快速且可逆的扰动。
