Visualizing the distribution and transport of mRNAs in living cells
Visualizing the distribution and transport of mRNAs in living cells
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DOI：
10.1073/pnas.2233244100
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发表时间：
2003-11-11
影响因子：
11.1
通讯作者：
Tyagi, S
中科院分区：
文献类型：
--
作者：
Bratu, DP;Cha, BJ;Tyagi, S
我们已经观察到了活细胞中天然信使核糖核酸（mRNAs）的运动。利用抗核酸酶的分子信标，我们对黑腹果蝇卵母细胞中奥斯卡（oskar）mRNA的运输和定位进行了成像。当通过对mRNA的3'非翻译区进行基因操作，或者通过对细胞内微管蛋白网络进行化学扰动来改变定位模式时，荧光信号的分布也会相应地改变。我们实时追踪了奥斯卡mRNA从产生它的滋养细胞到它所定位的卵母细胞后皮质的迁移过程。我们的观察结果揭示了奥斯卡mRNA运输过程中一个此前难以捉摸的短暂阶段的存在。利用分子信标对活细胞中特定的mRNAs进行直接观察，将加速细胞内RNA运输和定位的研究，就像绿色荧光蛋白的使用促进了体内特定蛋白质的研究一样。
We have visualized the movements of native mRNAs in living cells. Using nuclease-resistant molecular beacons, we imaged the transport and localization of oskar mRNA in Drosophila melanogaster oocytes. When the localization pattern was altered by genetic manipulation of the mRNA's 3' untranslated region, or by chemical perturbation of the intracellular tubulin network, the distribution of the fluorescence signals changed accordingly. We tracked the migration of oskar mRNA in real time, from the nurse cells where it is produced to the posterior cortex of the oocyte where it is localized. Our observations reveal the presence of a transient, and heretofore elusive, stage in the transport of oskar mRNA. Direct visualization of specific mRNAs in living cells with molecular beacons will accelerate studies of intracellular RNA trafficking and localization, just as the use of green fluorescent protein has stimulated the study of specific proteins in vivo.