Inducible control of subcellular RNA localization using a synthetic protein-RNA aptamer interaction.
Inducible control of subcellular RNA localization using a synthetic protein-RNA aptamer interaction.
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DOI：
10.1371/journal.pone.0046868
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Niles JC
中科院分区：
文献类型：
--
作者：
Belmont BJ;Niles JC
越来越多的证据支持亚细胞RNA定位在不同生物学环境中的功能重要性。在不同的细胞类型中，经常观察到不同的RNA定位模式，现有数据表明这是通过一系列高度协调的事件实现的。传统上，待定位的RNA内的顺式元件被RNA结合蛋白（RBP）识别，然后指导靶RNA的特异性定位。到目前为止，精确控制的时空参数固有的调节RNA的本地化还没有实验上是可能的。在这里，我们展示了化学诱导的RNA-蛋白质相互作用来调节亚细胞RNA定位的发展和使用。我们的系统主要由两部分组成：（i）与天然参与定位内源性转录物的蛋白质遗传融合的泰特阻遏蛋白（TetR）;和（ii）含有遗传编码的TetR结合RNA适体的靶转录物。TetR融合蛋白与靶RNA的结合以及后者的随后定位直接受多西环素调节。使用这个平台，我们证明，增强和控制的亚细胞定位的工程转录本是可以实现的。我们还分析了正向工程的RNA定位系统的规则，以促进其直接的应用，更普遍地研究RNA定位。
Evidence is accumulating in support of the functional importance of subcellular RNA localization in diverse biological contexts. In different cell types, distinct RNA localization patterns are frequently observed, and the available data indicate that this is achieved through a series of highly coordinated events. Classically, cis–elements within the RNA to be localized are recognized by RNA-binding proteins (RBPs), which then direct specific localization of a target RNA. Until now, the precise control of the spatiotemporal parameters inherent to regulating RNA localization has not been experimentally possible. Here, we demonstrate the development and use of a chemically–inducible RNA–protein interaction to regulate subcellular RNA localization. Our system is composed primarily of two parts: (i) the Tet Repressor protein (TetR) genetically fused to proteins natively involved in localizing endogenous transcripts; and (ii) a target transcript containing genetically encoded TetR–binding RNA aptamers. TetR–fusion protein binding to the target RNA and subsequent localization of the latter are directly regulated by doxycycline. Using this platform, we demonstrate that enhanced and controlled subcellular localization of engineered transcripts are achievable. We also analyze rules for forward engineering this RNA localization system in an effort to facilitate its straightforward application to studying RNA localization more generally.
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影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
10.5
作者：
Gu, W;Deng, YF;Singer, RH
通讯作者：
Singer, RH
影响因子：
9.2
作者：
Beach, DL;Salmon, ED;Bloom, K
通讯作者：
Bloom, K
DOI：
10.1083/jcb.200705163
发表时间：
2007-12-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Blower MD;Feric E;Weis K;Heald R
通讯作者：
Heald R
影响因子：
64.5
作者：
Lecuyer, Eric;Yoshida, Hideki;Krause, Henry M.
通讯作者：
Krause, Henry M.