Use of RNA interference to dissect the roles of trans-acting factors in alternative pre-mRNA splicing.
Use of RNA interference to dissect the roles of trans-acting factors in alternative pre-mRNA splicing.
复制标题
DOI：
10.1016/j.ymeth.2005.07.016
复制
发表时间：
2005-12
期刊：
影响因子：
4.8
通讯作者：
J. Park;B. Graveley
中科院分区：
文献类型：
--
作者：
J. Park;B. Graveley
RNA干扰(RNAi)正成为分析各种生物过程中基因功能的一种流行方法。我们在培养的果蝇细胞中使用RNAi来识别调节内源性转录前mRNAs选择性剪接的反式作用因子。我们已经建立了一个双链RNA文库，该文库包含了编码∼结合蛋白的70%的果蝇基因，并评估了每种蛋白在调节选择性剪接中的功能。这种方法不仅可以识别调节特定选择性剪接事件的反式作用因子，而且可以为深入了解果蝇的选择性剪接调控网络提供依据。在这里，我们详细描述了这种RNAi方法来识别替代剪接调节蛋白。
RNA interference (RNAi) is becoming a popular method for analyzing gene function in a variety of biological processes. We have used RNAi in cultured Drosophila cells to identify trans-acting factors that regulate the alternative splicing of endogenously transcribed pre-mRNAs. We have generated a dsRNA library comprising ∼70% of the Drosophila genes encoding RNA binding proteins and assessed the function of each protein in the regulation of alternative splicing. This approach not only identifies trans-acting factors regulating specific alternative splicing events, but also can provide insight into the alternative splicing regulatory networks of Drosophila. Here, we describe this RNAi approach to identify alternative splicing regulatory proteins in detail.