Functional siRNA expression from transfected PCR products
Functional siRNA expression from transfected PCR products
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DOI：
10.1017/s1355838202021362
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发表时间：
2002-11-01
期刊：
影响因子：
4.5
通讯作者：
Rossi, JJ
中科院分区：
文献类型：
--
作者：
Castanotto, D;Li, HT;Rossi, JJ
RNA 干扰 (RNAi) 是双链 RNA (dsRNA) 诱导同源转录物转录后降解的过程。 RNAi 可以通过转染或内源表达将细胞暴露于 dsRNA 来启动。在哺乳动物系统中，通过能够形成双链体的21-23碱基转录物的表达，或通过短发夹RNA的表达，观察到了序列特异性RNAi效应。我们在这里描述了一种基于 PCR 的简单策略，用于快速合成 siRNA 表达单元及其在哺乳动物细胞中的测试。通过PCR构建siRNA表达构建体，并将PCR产物直接转染至哺乳动物细胞中，从而实现siRNA的功能性表达。这种方法对于鉴定最佳 siRNA-靶标组合以及在哺乳动物细胞中多重 siRNA 表达应该是有用的。
RNA interference (RNAi) is a process in which double-stranded RNA (dsRNA) induces the postranscriptional degradation of homologous transcripts. RNAi can be initiated by exposing cells to dsRNA either via transfection or endogenous expression. In mammalian systems, the sequence-specific RNAi effect has been observed by expression of 21-23 base transcripts capable of forming duplexes, or via expression of short hairpin RNAs. We describe here a facile PCR based strategy for rapid synthesis of siRNA expression units and their testing in mammalian cells. The siRNA expression constructs are constructed by PCR, and the PCR products are directly transfected into mammalian cells resulting in functional expression of siRNAs. This approach should prove useful for identification of optimal siRNA-target combinations and for multiplexing siRNA expression in mammalian cells.