Tissue-specific and reversible RNA interference in transgenic mice
Tissue-specific and reversible RNA interference in transgenic mice
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DOI：
10.1038/ng2045
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发表时间：
2007-07-01
期刊：
影响因子：
30.8
通讯作者：
Lowe, Scott W.
中科院分区：
文献类型：
--
作者：
Dickins, Ross A.;McJunkin, Katherine;Lowe, Scott W.
基因工程小鼠为了解哺乳动物的基因功能提供了强大的工具。这些模型传统上依赖于转基因或靶向的、不可逆的基因突变导致的基因过度表达。通过采用以前用于基因过度表达的四环素(TET)反应系统，我们开发了一种简单的转基因系统，利用RNA干扰(RNAi)在小鼠中可逆地控制内源基因的表达。携带TET反应RNA聚合酶II启动子的转基因小鼠，当与表达一般或组织特异性‘tet-on’或‘tet-off’反式激活剂的现有小鼠品系杂交时，可可逆地表达针对肿瘤抑制基因Trp53的基于microRNA的短发夹RNA。TrP53基因敲除可在多个组织中实现可逆性下调，在淋巴瘤中恢复TrP53的表达可导致肿瘤的消退。通过保持靶基因不变，这种方法允许在体内对内源性基因表达进行组织特异性的可逆调节，在基础生物学和药物靶点验证方面具有潜在的广泛应用前景。
Genetically engineered mice provide powerful tools for understanding mammalian gene function. These models traditionally rely on gene overexpression from transgenes or targeted, irreversible gene mutation. By adapting the tetracycline ( tet)- responsive system previously used for gene overexpression, we have developed a simple transgenic system to reversibly control endogenous gene expression using RNA interference ( RNAi) in mice. Transgenic mice harboring a tet- responsive RNA polymerase II promoter driving a microRNA- based short hairpin RNA targeting the tumor suppressor Trp53 reversibly express short hairpin RNA when crossed with existing mouse strains expressing general or tissue- specific 'tet- on' or 'tet- off' transactivators. Reversible Trp53 knockdown can be achieved in several tissues, and restoring Trp53 expression in lymphomas whose development is promoted by Trp53 knockdown leads to tumor regression. By leaving the target gene unaltered, this approach permits tissue- specific, reversible regulation of endogenous gene expression in vivo, with potential broad application in basic biology and drug target validation.