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ACS Synth Biol. 2016 Mar 18;5(3):207-12. doi: 10.1021/acssynbio.5b00264. Epub 2015 Dec 28.

Multiple Gene Repression in Cyanobacteria Using CRISPRi.

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KTH-Royal Institute of Technology , Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, Stockholm SE-171 21 Sweden.


We describe the application of clustered regularly interspaced short palindromic repeats interference (CRISPRi) for gene repression in the model cyanobacterium Synechcocystis sp. PCC 6803. The nuclease-deficient Cas9 from the type-II CRISPR/Cas of Streptrococcus pyogenes was used to repress green fluorescent protein (GFP) to negligible levels. CRISPRi was also used to repress formation of carbon storage compounds polyhydroxybutryate (PHB) and glycogen during nitrogen starvation. As an example of the potential of CRISPRi for basic and applied cyanobacteria research, we simultaneously knocked down 4 putative aldehyde reductases and dehydrogenases at 50-95% repression. This work also demonstrates that tightly repressed promoters allow for inducible and reversible CRISPRi in cyanobacteria.

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