期刊
ACS SYNTHETIC BIOLOGY
卷 5, 期 1, 页码 74-80出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.5b00130
关键词
translational activator; Csy4; CRISPR endoribonuclease; modularization; RBS calculator; orthogonal parts
资金
- Ministry of Science and Technology of China (973 Grant) [2013CB734001]
- National Natural Science Foundation of China [31470818]
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.
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