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论文题目: Engineering Translational Activators with CRISPR-Cas System
作者: Du Pei , Miao Chensi , Lou Qiuli , Wang Zefeng* , and Lou Chunbo*.
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刊物名称: Acs Synthetic Biology
期: 1
卷: 5
页: 74-80
年份: 2016
影响因子: 5.037
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摘要: RNA parts often serve as. critical components in genetic engineering. Here we report a design of translational activators which is composed of an RNA endoribonuclease (Csy4) and two exchaneable RNA modules. Csy4, member of Cas endoribonuclease, cleaves at a specific recognition site; this cleavage releases a cis-repressive RNA module (crRNA) from the masked ribosome binding site (RBS), which subsequently allows the downstream translation initiation. Unlike small RNA as a translational activator, the endoribonuclease-based activator is able, to efficiently unfold the perfect RBS-crRNA pairing. As an exchangeable module, the crRNA-RBS duplex was forwardly and reversely engineered to modulate the dynamic range of translational activity. We further showed that Csy4 and its recognition site, together as a module, can also be replaced by orthogonal endoribonuclease-recognition site homologues. These modularly structured, high-performance translational activators would endow the programming of gene expression in, the translation level with higher feasibility.