Abstract
Cyanobacteria are an attractive host for biofuel production because they can produce valuable chemical compounds from CO2 fixed by photosynthesis. However, the available genetic tools that enable precise gene regulation for the applications of synthetic biology are insufficient. Previously, we engineered an RNA-based posttranscriptional regulator, termed riboregulator, for the control of target gene expression in cyanobacterium Synechocystis sp. PCC 6803. Moreover, we enhanced the gene regulation ability of the riboregulators in Escherichia coli by fusing and engineering a scaffold sequence derived from naturally occurring E. coli noncoding small RNAs. Here, we demonstrated that the scaffold sequence fused to the riboregulators improved their gene regulation ability in Synechocystis sp. PCC 6803. To further improve gene regulation, we expressed an exogenous RNA chaperone protein that is responsible for noncoding small RNA-mediated gene regulation, which resulted in higher target gene expression. The scaffold sequence derived from natural E. coli noncoding small RNAs is effective for designing RNA-based genetic tools and scaffold-fused riboregulators are a strong RNA-tool to regulate gene expression in cyanobacteria.
Highlights
Cyanobacteria have great potential as a host for biofuel production owing to their photosynthesis ability, higher growth rate than plants, and the ease of genetic engineering
While diverse and powerful gene regulators have been designed for E. coli, the genetic tools engineered in cyanobacteria are limited to only a few inducible promoters (Huang et al 2010; Huang and Lindblad 2013; Abe et al 2014a; Camsund et al 2014) or RNA-based tools (Nakahira et al 2013; Abe et al 2014b)
The scaffoldfused taR*2’s were composed of taR*2, a previously engineered riboregulator suitable for gene regulation in Synechocystis, and the small RNA (sRNA) scaffold sequence derived from natural E. coli MicF sRNA
Summary
Cyanobacteria have great potential as a host for biofuel production owing to their photosynthesis ability, higher growth rate than plants, and the ease of genetic engineering. They fix CO2 from air by photosynthesis and convert it into chemical products via biosynthetic pathways (Niederholtmeyer et al 2010; Oliver et al 2013; Osanai et al 2013). While diverse and powerful gene regulators have been designed for E. coli, the genetic tools engineered in cyanobacteria are limited to only a few inducible promoters (Huang et al 2010; Huang and Lindblad 2013; Abe et al 2014a; Camsund et al 2014) or RNA-based tools (Nakahira et al 2013; Abe et al 2014b). To expand the synthetic biology applications using cyanobacteria, more diverse and powerful gene regulators are required
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