Temporal dissection of p53 function in vitro and in vivo
Temporal dissection of p53 function in vitro and in vivo
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DOI：
10.1038/ng1572
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发表时间：
2005-07-01
期刊：
影响因子：
30.8
通讯作者：
Evan, GI
中科院分区：
文献类型：
--
作者：
Christophorou, MA;Martin-Zanca, D;Evan, GI
为了研究p53肿瘤抑制因子的功能，我们建立了一种新的敲入基因置换小鼠模型，其中内源性Trp 53基因被编码p53 ER（TAM）的基因置换，p53 ER（TAM）是一种p53融合蛋白，其功能完全依赖于4-羟基他莫昔芬的异位提供。我们在这里表明，来自这些小鼠的体内组织和体外细胞都可以在野生型和p53敲除状态之间快速切换。使用这种快速扰动模型，我们定义的动力学，依赖性，持久性和可逆性的p53介导的反应，在体内组织中的DNA损伤和激活的Ras癌蛋白和应力在体外。这是我们所知的一类新的遗传模型的第一个例子，该模型允许对体内单个内源基因的功能进行特异性、快速和可逆的扰动。
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.