An epi-allelic series of p53 hypomorphs created by stable RNAi produces distinct tumor phenotypes in vivo
An epi-allelic series of p53 hypomorphs created by stable RNAi produces distinct tumor phenotypes in vivo
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DOI：
10.1038/ng1091
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发表时间：
2003-03-01
期刊：
影响因子：
30.8
通讯作者：
Lowe, SW
中科院分区：
文献类型：
--
作者：
Hemann, MT;Fridman, JS;Lowe, SW
RNA干扰（RNAi）在哺乳动物系统中的应用有可能彻底改变遗传学并产生新的治疗方法。在这里，我们调查是否RNAi应用于一个良好的特点基因可以稳定地抑制造血干细胞的基因表达，并产生可检测的表型在小鼠。Trp 53肿瘤抑制基因的缺失大大加速Myc诱导的淋巴瘤发生，导致高度播散性疾病（1，2）。为了确定Trp 53的RNAi抑制是否可以产生类似的表型，我们将几种Trp 53短发夹RNA（shRNAs）引入来自Emu-Myc转基因小鼠的造血干细胞中，并监测致死辐射受体的肿瘤发作和总体病理学。不同的Trp 53 shRNA在体内产生不同的表型，从良性淋巴增生到高度播散性淋巴瘤，这些淋巴瘤在Emu-Myc小鼠中抑制Trp 53（-/-）淋巴瘤发生。在所有情况下，疾病的严重程度和类型与特定shRNA抑制p53活性的程度相关。因此，RNAi可以稳定地抑制干细胞和源自这些细胞的重建器官中的基因表达。此外，靶向相同基因的单个shRNA表达载体之间的内在差异可用于创建用于解剖体内基因功能的“表观等位基因系列”。
The application of RNA interference (RNAi) to mammalian systems has the potential to revolutionize genetics and produce novel therapies. Here we investigate whether RNAi applied to a well-characterized gene can stably suppress gene expression in hematopoietic stem cells and produce detectable phenotypes in mice. Deletion of the Trp53 tumor suppressor gene greatly accelerates Myc-induced lymphomagenesis, resulting in highly disseminated disease(1,2). To determine whether RNAi suppression of Trp53 could produce a similar phenotype, we introduced several Trp53 short hairpin RNAs (shRNAs) into hematopoietic stem cells derived from Emu-Myc transgenic mice, and monitored tumor onset and overall pathology in lethally irradiated recipients. Different Trp53 shRNAs produced distinct phenotypes in vivo, ranging from benign lymphoid hyperplasias to highly disseminated lymphomas that paralleled Trp53(-/-) lymphomagenesis in the Emu-Myc mouse. In all cases, the severity and type of disease correlated with the extent to which specific shRNAs inhibited p53 activity. Therefore, RNAi can stably suppress gene expression in stem cells and reconstituted organs derived from those cells. In addition, intrinsic differences between individual shRNA expression vectors targeting the same gene can be used to create an 'epi-allelic series' for dissecting gene function in vivo.