ACS photonics, 2016-03, Vol.5 (3), p.207-212
2016
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- Autor(en) / Beteiligte
- Titel
- Multiple Gene Repression in Cyanobacteria Using CRISPRi
- Ist Teil von
- ACS photonics, 2016-03, Vol.5 (3), p.207-212
- Ort / Verlag
- United States: American Chemical Society
- Erscheinungsjahr
- 2016
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2161-5063, 2330-4022eISSN: 2161-5063, 2330-4022DOI: 10.1021/acssynbio.5b00264Titel-ID: cdi_swepub_primary_oai_DiVA_org_kth_185366
- Format
- –
- Schlagworte
- Aldehyde, Aldehyde Dehydrogenase - genetics, Aldehyde Dehydrogenase - metabolism, Aldehyde Reductase - genetics, Aldehyde Reductase - metabolism, Bacterial Proteins - genetics, Bacterial Proteins - metabolism, Blotting, Western, Carbon-Dioxide, CRISPR-Cas Systems - genetics, Cyanobacteria - metabolism, Dehydrogenase, Down-Regulation, Escherichia-Coli, Expression, Flow Cytometry, Green Fluorescent Proteins - genetics, Green Fluorescent Proteins - metabolism, Light, Sequence-Specific Control, Strain Pcc 6803, Synechocystis Sp Pcc6803, Tolerance