ArticleMicrobial cell factories2016
CRISPR interference (CRISPRi) for gene regulation and succinate production in cyanobacterium S. elongatus PCC 7942.
Article in Microbial cell factories, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed, 162 citations in OpenAlex.
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- dCas12a-mediated CRISPR interference for multiplex gene repression in cyanobacteria for enhanced isobutanol and 3-methyl-1-butanol production.Microbial cell factories · 2025Article
- The Strategy and Application of Gene Attenuation in Metabolic Engineering.Microorganisms · 2025Review
- Cyanobacterial type I CRISPR-Cas systems: distribution, mechanisms, and genome editing applications.Frontiers in bioengineering and biotechnology · 2025Review
- Targeting Transcriptional Regulators Affecting Acarbose Biosynthesis inMicroorganisms · 2024Article
- Insufficient Acetyl-CoA Pool Restricts the Phototrophic Production of Organic Acids in Model Cyanobacteria.International journal of molecular sciences · 2024Review
- Improving Lipid Content in the DiatomCurrent issues in molecular biology · 2024Article
- CRISPRi functional genomics in bacteria and its application to medical and industrial research.Microbiology and molecular biology reviews : MMBR · 2024Review
- A potential paradigm in CRISPR/Cas systems delivery: at the crossroad of microalgal gene editing and algal-mediated nanoparticles.Journal of nanobiotechnology · 2023Review
- Genome-scale CRISPRi screening: A powerful tool in engineering microbiology.Engineering microbiology · 2023Review
- Cyanobacteria as cell factories for the photosynthetic production of sucrose.Frontiers in microbiology · 2023Review
- Transcription factor-based biosensors for screening and dynamic regulation.Frontiers in bioengineering and biotechnology · 2023Review
- Metabolic Engineering Design Strategies for Increasing Carbon Fluxes Relevant for Biosynthesis in Cyanobacteria.Advances in biochemical engineering/biotechnology · 2023Article
- Activating natural product synthesis using CRISPR interference and activation systems in Streptomyces.Nucleic acids research · 2022Article
- Combinatorial CRISPR Interference Library for Enhancing 2,3-BDO Production and Elucidating Key Genes in Cyanobacteria.Frontiers in bioengineering and biotechnology · 2022Article
- CRISPR interference for sequence-specific regulation of fibroblast growth factor receptor A inFrontiers in immunology · 2022Article
- CRISPR-Based Approaches for Gene Regulation in Non-Model Bacteria.Frontiers in genome editing · 2022Review
- Comprehensive Genome Engineering Toolbox for MicroalgaeACS synthetic biology · 2021Article
- Application of CRISPR/Cas System in the Metabolic Engineering of Small Molecules.Molecular biotechnology · 2021Review
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundCyanobacterium Synechococcus elongatus PCC 7942 holds promise for biochemical conversion, but gene deletion in PCC 7942 is time-consuming and may be lethal to cells. CRISPR interference (CRISPRi) is an emerging technology that exploits the catalytically inactive Cas9 (dCas9) and single guide RNA (sgRNA) to repress sequence-specific genes without the need of gene knockout, and is repurposed to rewire metabolic networks in various procaryotic cells.
resultsTo employ CRISPRi for the manipulation of gene network in PCC 7942, we integrated the cassettes expressing enhanced yellow fluorescent protein (EYFP), dCas9 and sgRNA targeting different regions on eyfp into the PCC 7942 chromosome. Co-expression of dCas9 and sgRNA conferred effective and stable suppression of EYFP production at efficiencies exceeding 99%, without impairing cell growth. We next integrated the dCas9 and sgRNA targeting endogenous genes essential for glycogen accumulation (glgc) and succinate conversion to fumarate (sdhA and sdhB). Transcription levels of glgc, sdhA and sdhB were effectively suppressed with efficiencies depending on the sgRNA binding site. Targeted suppression of glgc reduced the expression to 6.2%, attenuated the glycogen accumulation to 4.8% and significantly enhanced the succinate titer. Targeting sdhA or sdhB also effectively downregulated the gene expression and enhanced the succinate titer ≈12.5-fold to ≈0.58-0.63 mg/L.
conclusionsThese data demonstrated that CRISPRi-mediated gene suppression allowed for re-directing the cellular carbon flow, thus paving a new avenue to rationally fine-tune the metabolic pathways in PCC 7942 for the production of biotechnological products.
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