Heritable and stable gene knockdown in rats
Heritable and stable gene knockdown in rats
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DOI：
10.1073/pnas.0604657103
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发表时间：
2006-07-25
影响因子：
11.1
通讯作者：
Garbers, David L.
中科院分区：
文献类型：
--
作者：
Dann, Christina Tenenhaus;Alvarado, Alma L.;Garbers, David L.
大鼠一直是动物生理学研究的极好模型，也是糖尿病和酒精中毒等人类疾病的模型；然而，由于无法敲除基因，遗传研究一直受到限制。我们的目标是在大鼠体内利用RNA干扰来抑制基因表达，从而在特定基因功能上产生可遗传的缺陷。慢病毒介导的转基因被用来产生针对DAZL的短发夹状RNA，DAZL是一种在生殖细胞中表达的基因，是小鼠生育所必需的基因[Ruggiu，M.，Speed，R.，Taggart，M.，McKay，S.J.，Kilanowski，F.，Saunders，P.，Dorin，J.&Cooke，H.J.(1997)Nature 389，73-77]。转基因的生殖系传递发生了，其表达与DAZL蛋白水平的显著降低和雄性不育有关，并且在多个世代(F-1-F-3)中敲除是稳定的。这项研究展示了一种有效的系统，通过该系统，现在可以在大鼠身上进行定向反向遗传分析。
The rat has served as an excellent model for studies on animal physiology and as a model for human diseases such as diabetes and alcoholism; however, genetic studies have been limited because of the inability to knock out genes. Our goal was to produce heritable deficiencies in specific gene function in the rat using RNA interference to knock down gene expression in vivo. Lentiviral-mediated transgenesis was used to produce rats expressing a short hairpin RNA targeting Dazl, a gene expressed in germ cells and required for fertility in mice [Ruggiu, M., Speed, R., Taggart, M., McKay, S. J., Kilanowski, F., Saunders, P., Dorin, J. & Cooke, H. J. (1997) Nature 389, 73-77]. Germ-line transmission of the transgene occurred, and its expression correlated with significant reductions in DAZL protein levels and male sterility, and the knockdown was stable over multiple generations (F-1-F-3). This study demonstrates an efficient system by which directed reverse genetic analysis can now be performed in the rat.