The artificial microRNA mediates GUS-GFP gene silencing using ath-miR169d precursor as backbone

文献类型： 外文期刊

第一作者： Liu, Chong

作者： Liu, Chong;Zhang, Lan;Sun, Jie;Luo, Yanzhong;Fan, Yunliu;Wang, Lei;Liu, Chong;Sun, Jie;Wang, Mingbo

作者机构：

关键词： miRNA;hairpin RNA;artificial microRNA;silencing gene

期刊名称：LIFE SCIENCE JOURNAL-ACTA ZHENGZHOU UNIVERSITY OVERSEAS EDITION （ 影响因子：0.165； ）

ISSN： 1097-8135

年卷期： 2009 年 6 卷 2 期

页码：

收录情况： SCI

摘要： Artificial microRNA (amiRNA) is becoming a powerful tool for silencing genes in plants, and several amiRNA vectors have recently been developed based on the natural precursor structures of ath-miR159a, ath-miR164b, ath-miR172a, ath-miR319a and osa-miR528. In this study we generated a simple amiRNA vector (pAmiR169d) based on the structure of Arabidopsis miR169d precursor (pre-miR169d). Two unique restriction sites were created inside the stem region of pre-miR169d, which allows for amiRNA sequences to be cloned as either similar to 80 bp synthetic oligonucleotides or PCR products. A beta-glucuronidase (GUS)/green florescent protein (GFP) fusion gene was efficiently silenced in transient assays using a pAmiR169d-derived construct targeting a GFP sequence. 5' RACE showed that the target GFP transcript was cleaved precisely at the expected position across nucleotides 10 and 11 of the amiRNA. Thus, pAmiR169d allows for both easy construction of amiRNA constructs and efficient silencing of target genes in plants. [Life Science Journal. 2009; 6(2): 1 - 7] (ISSN: 1097 - 8135).

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