摘要
在哺乳动物的RNAi研究中,载体表达的shRNA分子比细胞同时表达的siRNA分子的正义链与反义链对靶基因的抑制效率要高。shRNA可由PolⅢ的启动子在体内表达产生,酶切cDNA和shRNA芯片是产生shRNA的最新方法。对shRNA的设计应注意靶基因序列、环序列以及载体酶切位点的选择。诱导表达shRNA的载体系统的表达效率有所差异,质粒载体转染效率尚不稳定,且持续时间短,通过病毒载体介导是目前进行基因敲除最有效的工具。
In mammlian RNAi,vector-based short interfering RNA（siRNA） is usually generated through short hairpin RNA （shRNA）. In this system, RNA Pol Ⅲ promoters are used to drive transcription of shRNA. Alternative approach to generate shRNA from the target cDNA by enzymatic digest and Parallel synthesis of thousands of oligonucleotides in situ on microarmayy plaffomas. The shRNA- mediated gene silencing efficiency is determined by many factors;some major factors are the selection of target, loop sequences and enzyme site. The major advantage of vector-based RNAi is the achievement of long-lasting gene silencing with shRNAs expressed from stably transfected plasmids or from integrated retroor lentiviral vectors. This review will focus on the practical aspects of shRNA conducted in mammalian RNAi systems, emphasizing the design, delivery, functional validation of targeting molecules and inducible system.
出处
《生物技术》
CAS
CSCD
2006年第2期79-82,共4页
Biotechnology
