Primer extension-based method for the generation of a siRNA/miRNA expression vector
Primer extension-based method for the generation of a siRNA/miRNA expression vector
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DOI：
10.1152/physiolgenomics.00005.2007
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发表时间：
2007-11-14
影响因子：
4.6
通讯作者：
Liu, Lin
中科院分区：
文献类型：
--
作者：
Gou, Deming;Zhang, Honghao;Liu, Lin
RNA干扰（RNAi）已成为研究基因功能、生物学途径和疾病生理学的有力技术。通常，哺乳动物细胞中的RNAi应答由小干扰RNA（siRNA）介导。使用合成的siRNA沉默基因是相对快速和容易的，但它是昂贵的瞬时效应。具有由环结构分开的靶基因的互补正义和反义序列的短发夹RNA（shRNA）导致基因沉默，其与化学合成的siRNA一样有效，具有更少的限制。然而，目前构建shRNA载体的方法需要合成长寡核苷酸，成本高昂，并且在合成过程中经常遇到突变问题。在这里，我们报告了一种替代方法，以产生一个高效的shRNA表达载体。我们使用较短的（
RNA interference (RNAi) has become a powerful technique for studying gene function, biological pathways, and the physiology of diseases. Typically, the RNAi response in mammalian cells is mediated by small interfering RNA (siRNA). The use of synthesized siRNA to silence gene is relatively quick and easy, but it is costly with transient effects. A short hairpin RNA (shRNA) with complementary sense and antisense sequences of a target gene separated by a loop structure results in gene silencing that is as effective as chemically synthesized siRNA with fewer limitations. However, current methods for constructing shRNA vectors require the synthesis of long oligonucleotides, which is costly and often suffers from mutation problems during synthesis. Here, we report an alternative approach to generate a shRNA expression vector with high efficacy. We utilized shorter (