Multiple Gene Repression in Cyanobacteria Using CRISPRi
2016 (English)In: ACS Photonics, ISSN 2186-2311, E-ISSN 2161-5063, Vol. 5, no 3, 207-212 p.Article in journal (Refereed) PublishedText
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.
Place, publisher, year, edition, pages
American Chemical Society (ACS), 2016. Vol. 5, no 3, 207-212 p.
Strain Pcc 6803, Sequence-Specific Control, Synechocystis Sp Pcc6803, Escherichia-Coli, Carbon-Dioxide, Light, Expression, Dehydrogenase, Tolerance, Aldehyde
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-185366DOI: 10.1021/acssynbio.5b00264ISI: 000372672500003PubMedID: 26689101ScopusID: 2-s2.0-84961794371OAI: oai:DiVA.org:kth-185366DiVA: diva2:920613
FunderSwedish Research Council Formas, 213-2011-1655Science for Life Laboratory - a national resource center for high-throughput molecular bioscienceSwedish Foundation for Strategic Research , RBP14-0013
QC 201604182016-04-182016-04-182016-04-22Bibliographically approved