由抑制性微RNA（miRNA）誘導的基因沉默在調(diào)控基因表達中的重要性現(xiàn)已非常明顯,。但因為一個信使RNA（mRNA）的miRNA調(diào)控在它們的序列中只需要有一個短的匹配段（長度為8個核苷酸或更少）,，所以事實證明,，要明確確定所預測的很多mRNA結(jié)合點中哪個是每個miRNA在活體中的目標幾乎不可能,。
現(xiàn)在,，對HITS-CLIP方法所做的一種改進（這種改進的方法以miRNA和 mRNA與Argonaute蛋白的相互作用為關(guān)注點,；Argonaute蛋白是普遍存在的核酸內(nèi)切酶,，它們形成由RNA誘導的沉默復合物的一部分）,，被用來解析miRNA與大腦mRNA轉(zhuǎn)錄體的結(jié)合點的一個精確分布圖。該方法具有普遍適用性,，應能為了解miRNA在生物學中的作用提供了一個新手段,。
另外，這種分布圖可讓研究人員確定具有臨床意義的mRNA上用于RNA干涉（RNAi）療法的目標作用點,。（生物谷Bioon.com）
生物谷推薦原始出處：
Nature 460, 479-486 (23 July 2009) | doi:10.1038/nature08170
Argonaute HITS-CLIP decodes microRNA–mRNA interaction maps
Sung Wook Chi1, Julie B. Zang1, Aldo Mele1 & Robert B. Darnell1
1 Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA
MicroRNAs (miRNAs) have critical roles in the regulation of gene expression; however, as miRNA activity requires base pairing with only 6-8 nucleotides of messenger RNA, predicting target mRNAs is a major challenge. Recently, high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation (HITS-CLIP) has identified functional protein–RNA interaction sites. Here we use HITS-CLIP to covalently crosslink native argonaute (Ago, also called Eif2c) protein–RNA complexes in mouse brain. This produced two simultaneous data sets—Ago–miRNA and Ago–mRNA binding sites—that were combined with bioinformatic analysis to identify interaction sites between miRNA and target mRNA. We validated genome-wide interaction maps for miR-124, and generated additional maps for the 20 most abundant miRNAs present in P13 mouse brain. Ago HITS-CLIP provides a general platform for exploring the specificity and range of miRNA action in vivo, and identifies precise sequences for targeting clinically relevant miRNA–mRNA interactions.
