construction of an artificial microrna expression vector for simultaneous inhibition of multiple genes in mammalian cells建设人工微rna表达载体的同时抑制多个基因在哺乳动物细胞.pdf

Int. J. Mol. Sci. 2009, 10, 2158-2168; doi:10.3390/ijm OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms Communication Construction of an Artificial MicroRNA Expression Vector for Simultaneous Inhibition of Multiple Genes in Mammalian Cells Tao Hu 1, Qiong Fu 1, Ping Chen 2, Li Ma 1, Onsam Sin 1 and Deyin Guo 1,* 1 State Key Laboratory of Virology and The Modern Virology Research Centre, College of Life Sciences, Wuhan University, Wuhan 430072, China; E-Mails: hnhutao@ (T.H.); qiongfu@ (Q.F.); mar126.com (L.M.); soskorea@163.com (O.S.) 2 Department of Pathophysiology, Basic Medical College, Zhengzhou University, Zhengzhou 450001, China; E-Mail: chp7779@ (P.C.) * Author to whom correspondence should be addressed; E-Mail: dguo@; Tel. +86-27-6875-2506; Fax: +86-27-6875-2897 Received: 8 April 2009; in revised form: 16 April 2009 / Accepted: 14 May 2009 / Published: 14 May 2009 Abstract: Recently, artificial microRNA (amiRNA) has become a promising RNA interference (RNAi) technology. Here, we describe a flexible and reliable method for constructing both single- and multi-amiRNA expression vectors. Two universal primers, together with two specific primers carrying the encoding sequence of amiRNA were designed and utilized to synthesize the functional amiRNA cassette through a one-step PCR. With