Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells
Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells
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DOI：
10.1016/j.chembiol.2004.06.006
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发表时间：
2004-08-01
影响因子：
--
通讯作者：
Rana, TM
中科院分区：
文献类型：
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作者：
Chiu, YL;Ali, A;Rana, TM
RNA干扰（RNAi）是短干扰RNA （siRNA）通过与RNA诱导的沉默复合体（RISC）相互作用，靶向特定mRNA进行降解的过程。在这里，通过使用siRNA- tat（47-57）肽、siRNA- tat（47-57）衍生的寡氨基甲酸酯偶联物或纳米颗粒将siRNA递送到细胞中，发现了siRNA定位、细胞摄取和RNAi活性之间的明确相关性。对于成功的RNAi， siRNA的定位明显是在核周围，这表明siRNA靶向这些区域与RISC相互作用以诱导RNAi。siRNA序列的变化和靶mRNA的存在显然没有改变siRNA的亚细胞定位模式。有趣的是，与TAT（47-57）肽或TAT（47-57）衍生的低氨基甲酸酯偶联的siRNA导致了高效的RNAi活性和siRNA的核周定位，这与非偶联的游离TAT肽核核定位明显不同。这些结果表明，即使siRNA与TAT（47-57）肽结合，与RISC的相互作用也决定了siRNA的定位。
RNA interference (RNAi) is the process by which short-interfering RNA (siRNA) target a specific mRNA for degradation through interactions with an RNA-induced silencing complex (RISC). Here, a clear correlation between siRNA localization, cellular uptake, and RNAi activity was discovered by delivering siRNA into cells using siRNA-TAT(47-57) peptide, siRNA-TAT(47-57)-derived oligocarbamate conjugates, or nanoparticles. For successful RNAi, the localization of siRNA was distinctly perinuclear, suggesting that siRNA is targeted to these regions for interactions with RISC to induce RNAi. siRNA sequence variation and the presence of the target mRNA apparently did not change the subcellular localization pattern of siRNA. Intriguingly, siRNA conjugated to TAT(47-57) peptide or TAT(47-57)-derived oligocarbamate resulted in efficient RNAi activity and perinuclear localization of siRNA that was distinctly different from nonconjugated free TAT peptide nucleolar localization. These results suggest that interactions with RISC dictate siRNA localization even when siRNA is conjugated to TAT(47-57) peptide.