Detection and measurement of alternative splicing using splicing-sensitive microarrays
Detection and measurement of alternative splicing using splicing-sensitive microarrays
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DOI：
10.1016/j.ymeth.2005.09.007
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发表时间：
2005-12-01
期刊：
影响因子：
4.8
通讯作者：
Ares, M
中科院分区：
文献类型：
--
作者：
Srinivasan, K;Shiue, L;Ares, M
剪接和选择性剪接是后生动物生物遗传信息解释和表达的主要过程。剪接的研究正在从集中注意力的一组选定的范式选择性剪接事件的调控机制，剪接调控与基因组和细胞功能的全球整合的问题。因此，有必要采用并行方法来检测和测量选择性剪接。我们已经适应了剪接敏感的寡核苷酸微阵列用于估计剪接效率在酵母中的脊椎动物细胞和组织中的选择性剪接的研究。我们使用基因模型，结合剪接的知识，设计特异性的寡核苷酸来区分选择性剪接的mRNA。在这里，我们提出的主要策略，设计，应用和分析的斑点寡核苷酸阵列检测和测量的选择性剪接。我们证明了这些策略，使用两个内含子的酵母基因，已被改变，以产生不同数量的选择性剪接的RNA，以及通过分析选择性剪接在NCI 60癌细胞系。(c)2005年爱思唯尔公司All rights reserved.
Splicing and alternative splicing are major processes in the interpretation and expression of genetic information for metazoan organisms. The study of splicing is moving from focused attention on the regulatory mechanisms of a selected set of paradigmatic alternative splicing events to questions of global integration of splicing regulation with genome and cell function. For this reason, parallel methods for detecting and measuring alternative splicing are necessary. We have adapted the splicing-sensitive oligonucleotide microarrays used to estimate splicing efficiency in yeast to the study of alternative splicing in vertebrate cells and tissues. We use gene models incorporating knowledge about splicing to design oligonucleotides specific for discriminating alternatively spliced mRNAs from each other. Here we present the main strategies for design, application, and analysis of spotted oligonucleotide arrays for detection and measurement of alternative splicing. We demonstrate these strategies using a two-intron yeast gene that has been altered to produce different amounts of alternatively spliced RNAs, as well as by profiling alternative splicing in NCI 60 cancer cell lines. (c) 2005 Elsevier Inc. All rights reserved.