Enzymatic preparation of an artificial microRNA library
文献类型：期刊论文
| 作者 | Xue, L; Yuan, Q; Yang, Y; Wu, JR |
| 刊名 | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
 |
| 出版日期 | 2009 |
| 卷号 | 390期号:3页码:791-796 |
| 关键词 | MicroRNA AmiRNA library Enzymatic method Gene silencing |
| 通讯作者 | Wu, JR (reprint author), Chinese Acad Sci, Key Lab Syst Biol, State Key Lab Mol Biol,Grad Univ, Inst Biochem & Cell Biol,Shanghai Inst Biol Sci, 320 Yueyang Rd, Shanghai 200031, Peoples R China.,wujr@sibs.ac.cn |
| 英文摘要 | MicroRNAs (miRNAs) are endogenous RNAs of about 20-25 nucleotides that play important roles in gene regulation. Based on miRNA backbones, artificial miRNAs, amiRNAs, can be constructed and expressed exogenously. AmiRNAs can function as natural miRNAs or siRNAs. AmiRNAs display higher expression levels and more potent silencing efficiency than simple shRNAs. In practice, an amiRNA library is usually needed to screen for functional amiRNAs. Here we describe a novel, convenient, and cost-effective method for enzymatically constructing an amiRNA library. A novel strategy was applied to ensure the correct orientation of the anti-sense strand and also render less thermo-stability to the 5' end of antisense strand to facilitate its entry into the RISC complex, which can largely increase the silencing efficiency. Furthermore, a "Padlock probe" method was also incorporated to ensure the fidelity of the sequence and stem-loop structure. To test the feasibility of the method, an amiRNA library targeting mouse p53 ORF was constructed. Through screening of 82 amiRNAs in the library, 7 amiRNAs which displayed potent p53-silencing efficiency were identified and verified. This method also has the potential to construct an amiRNA library that targets a whole transcriptome for genome-wide RNAi screening, or a randomized amiRNA library to search for functional amiRNAs. (C) 2009 Elsevier Inc. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | CAENORHABDITIS-ELEGANS ; PADLOCK PROBES ; RNAI LIBRARIES ; MESSENGER-RNAS ; HUMAN-CELLS ; TRANSLATION ; EXPRESSION ; TARGETS ; DIFFERENTIATION ; IDENTIFICATION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000272516700081 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1155]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Xue, L,Yuan, Q,Yang, Y,et al. Enzymatic preparation of an artificial microRNA library[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2009,390(3):791-796. |
| APA | Xue, L,Yuan, Q,Yang, Y,&Wu, JR.(2009).Enzymatic preparation of an artificial microRNA library.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,390(3),791-796. |
| MLA | Xue, L,et al."Enzymatic preparation of an artificial microRNA library".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 390.3(2009):791-796. |
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