Highly efficient small interfering RNA delivery to primary mammalian neurons induces MicroRNA-like effects before mRNA degradation
Highly efficient small interfering RNA delivery to primary mammalian neurons induces MicroRNA-like effects before mRNA degradation
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DOI：
10.1523/jneurosci.3643-04.2004
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发表时间：
2004-11-10
影响因子：
5.3
通讯作者：
Troy, CM
中科院分区：
文献类型：
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作者：
Davidson, TJ;Harel, S;Troy, CM
由于缺乏在神经元中诱导RNA干扰的高效、无毒的方法，对神经元蛋白质功能的研究一直受到阻碍。在这里，我们表明，应用合成的小干扰RNA(SiRNA)连接到载体肽Penetratin1导致整个培养的原始哺乳动物海马神经元和交感神经元群体快速、高效地摄取siRNA。这种处理方法可以在数小时内将目标蛋白特异性击倒，而不会产生与转基因相关的毒性。与目前的方法不同，我们的技术可以在最小程度上干扰复杂的细胞网络的情况下，研究整个种群的蛋白质功能。使用这项技术，我们发现蛋白质敲除(6小时后明显)先于靶向信息的任何减少(24小时后明显)，这表明完美靶向siRNA对早期的翻译抑制。
The study of protein function in neurons has been hindered by the lack of highly efficient, nontoxic methods of inducing RNA interference in such cells. Here we show that application of synthetic small interfering RNA( siRNA) linked to the vector peptide Penetratin1 results in rapid, highly efficient uptake of siRNA by entire populations of cultured primary mammalian hippocampal and sympathetic neurons. This treatment leads to specific knock-down of targeted proteins within hours without the toxicity associated with transfection. In contrast to current methods, our technique permits study of protein function across entire populations with minimal disturbance of complex cellular networks. Using this technique, we found that protein knock-down ( evident after 6 hr) precedes any decrease in targeted message ( evident after 24 hr), suggesting an early, translational repression by perfectly targeted siRNAs.