Specificity of short interfering RNA determined through gene expression signatures
Specificity of short interfering RNA determined through gene expression signatures
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DOI：
10.1073/pnas.1131959100
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发表时间：
2003-05-27
影响因子：
11.1
通讯作者：
Fesik, SW
中科院分区：
文献类型：
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作者：
Semizarov, D;Frost, L;Fesik, SW
短干扰RNA（siRNA）被广泛用于研究基因功能，并作为验证药物靶点和治疗疾病的工具具有很大的前景。这些应用中的关键假设是siRNA对细胞的作用是特异性的，即，仅限于靶基因的特异性敲低。在这篇文章中，我们通过应用基因表达谱来表征siRNA的特异性。针对三种不同靶标的相同靶基因的不同区域设计了几种siRNA。通过使用DNA微阵列生成基因表达特征来比较它们对细胞的影响。当siRNA设计和转染条件被优化时，针对相同靶标的不同siRNA的特征显示出非常密切的相关性，而不同基因的特征显示出没有相关性。这些结果表明，siRNA是一种高度特异性的靶向基因敲除工具，建立siRNA介导的基因沉默作为一种可靠的方法，用于大规模筛选基因功能和药物靶标验证。
Short interfering RNA (siRNA) is widely used for studying gene function and holds great promise as a tool for validating drug targets and treating disease. A critical assumption in these applications is that the effect of siRNA on cells is specific, i.e., limited to the specific knockdown of the target gene. In this article, we characterize the specificity of siRNA by applying gene expression profiling. Several siRNAs were designed against different regions of the same target gene for three different targets. Their effects on cells were compared by using DNA microarrays to generate gene expression signatures. When the siRNA design and transfection conditions were optimized, the signatures for different siRNAs against the same target were shown to correlate very closely, whereas the signatures for different genes revealed no correlation. These results indicate that siRNA is a highly specific tool for targeted gene knockdown, establishing siRNA-mediated gene silencing as a reliable approach for large-scale screening of gene function and drug target validation.