RNA silencing of single and multiple members in a gene family of rice
RNA silencing of single and multiple members in a gene family of rice
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DOI：
10.1104/pp.105.063933
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发表时间：
2005-08-01
期刊：
影响因子：
7.4
通讯作者：
Shimamoto, K
中科院分区：
文献类型：
--
作者：
Miki, D;Itoh, R;Shimamoto, K
RNA沉默与倒置重复（IR）结构已被用于抑制基因表达在各种生物体。然而，在植物中描述的传递性RNA沉默效应可能会阻止对基因家族使用RNA沉默。在这里，我们发现，在水稻（Oryza sativa）中，传递性RNA沉默（双链RNA沿着目标mRNA扩散）发生在绿色荧光蛋白转基因上，而不发生在内源性植物烯去饱和酶基因上。我们将来自水稻OsRac基因家族的独特的3'非翻译区域的IR拷贝融合到一个强启动子上，并将其稳定地引入水稻中。OsRac基因家族的7个成员中的每一个都被其各自的IR结构特异性抑制。我们还研究了融合了多个3'非翻译区的IR构建体，结果表明，一个构建体有效抑制了OsRac基因家族的三个成员。利用OsRac基因家族两个成员的高度保守区域，我们也以不同的效率抑制了该基因家族所有成员的表达。这些结果表明，RNA沉默是水稻和其他植物基因家族功能分析的一种有用方法。
RNA silencing with inverted repeat (IR) constructs has been used to suppress gene expression in various organisms. However, the transitive RNA-silencing effect described in plants may preclude the use of RNA silencing for a gene family. Here, we show that, in rice ( Oryza sativa), transitive RNA silencing ( spreading of double-stranded RNA along the target mRNA) occurred with the green fluorescent protein transgene but not with the endogenous phytoene desaturase gene. We fused IR copies of unique 3' untranslated regions derived from the rice OsRac gene family to a strong promoter and stably introduced them into rice. Each of the seven members of the OsRac gene family was specifically suppressed by its respective IR construct. We also examined IR constructs in which multiple 3' untranslated regions were fused and showed that three members of the OsRac gene family were effectively suppressed by a single construct. Using highly conserved regions of the two members of the OsRac gene family, we also suppressed the expression of all members of the gene family with variable efficiencies. These results suggest that RNA silencing is a useful method for the functional analysis of gene families in rice and other plants.