建立自然殺手細胞淋巴癌及霍奇金病細胞株的microRNA資料庫

Abstract

MicroRNA (miRNA) 是一種有調節能力的小分子的RNA，作為基因轉錄後的調控者。由於在基因表現上扮演重要角色，研究基因體 (genome)中miRNA的表現，傳統方法常使用定量反轉錄聚合酶鏈式反應(qRT-PCR) 或微陣列 (array) 分析miRNA。在這篇論文中，我們利用一種新的技術來分析基因體中miRNA的表現。藉由高效液相色層分析法(HPLC)純化出不同細胞株之miRNA，分別定序每一個選殖株 (clone)，並藉由生物資訊的方法來比對。每一個資料庫中約30-40%的選殖株為已知可信的miRNA

MicroRNAs (miRNAs) are a class of small regulatory RNAs, which act as post-transcriptional modulators of gene expression. Because of their critical roles in gene expression, genome-wide profiling of miRNAs, by qRT-PCR or array, has become an essential step in understanding cellular biology. In this thesis, we have developed a novel method for constructing miRNA libraries, as an initial step toward genome-wide miRNA characterization. We purified and cloned miRNAs by HPLC from several cell lines. Individual clones were sequenced, and identified by bioinformatics. It was estimated that about 30-40% of the clones from each library represented authentic miRNAs.