title: Multiple Gene Repression in Cyanobacteria Using CRISPRi.
abstract: 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.
authors.affiliation: KTH-Royal Institute of Technology , Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, Stockholm SE-171 21 Sweden.
authors.forename: Lun
authors.initials: L
authors.lastname: Yao
authors.affiliation: KTH-Royal Institute of Technology , Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, Stockholm SE-171 21 Sweden.
authors.forename: Ivana
authors.initials: I
authors.lastname: Cengic
authors.affiliation: KTH-Royal Institute of Technology , Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, Stockholm SE-171 21 Sweden.
authors.forename: Josefine
authors.initials: J
authors.lastname: Anfelt
authors.affiliation: KTH-Royal Institute of Technology , Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, Stockholm SE-171 21 Sweden.
authors.forename: Elton P
authors.initials: EP
authors.lastname: Hudson
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D064113:CRISPR-Cas Systems / Q000235:genetics*; D000458:Cyanobacteria / Q000378:metabolism*; D015536:Down-Regulation; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D049452:Green Fluorescent Proteins; D000449:Aldehyde Reductase; D000444:Aldehyde Dehydrogenase