ArticleMicrobial biotechnology2020
CRISPR interference (CRISPRi) as transcriptional repression tool for Hungateiclostridium thermocellum DSM 1313.
Article in Microbial biotechnology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 27 citations in OpenAlex.
- A robust enzymatic reporter system for the extremely thermophilic anaerobic bacteriumFrontiers in microbiology · 2026Article
- A fluorescent reporter system for trackingFrontiers in microbiology · 2026Article
- Designing thermophilic, synthetic microbial communities for consolidated bioprocessing.Biodesign research · 2025Review
- Biofuel production from lignocellulose via thermophile-based consolidated bioprocessing.Engineering microbiology · 2024Review
- CRISPRi functional genomics in bacteria and its application to medical and industrial research.Microbiology and molecular biology reviews : MMBR · 2024Review
- The past, present, and future promise of pluripotent stem cells.Journal of immunology and regenerative medicine · 2024Article
- The potential of native and engineered Clostridia for biomass biorefining.Frontiers in bioengineering and biotechnology · 2024Review
- A robust CRISPR interference gene repression system in Vibrio parahaemolyticus.Archives of microbiology · 2023Article
- Expression and Functional Analysis of the Compact ThermophilicInternational journal of molecular sciences · 2023Article
- Breaking the Restriction Barriers and Applying CRISPRi as a Gene Silencing Tool inMicroorganisms · 2022Article
- CRISPR-Based Approaches for Gene Regulation in Non-Model Bacteria.Frontiers in genome editing · 2022Review
- Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles.The CRISPR journal · 2021Article
- CRISPR interference (CRISPRi) as transcriptional repression tool for Hungateiclostridium thermocellum DSM 1313.Microbial biotechnology · 2020Article
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Authors and funding
3 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Hungateiclostridium thermocellum DSM 1313 has biotechnological potential as a whole-cell biocatalyst for ethanol production using lignocellulosic renewable sources. The full exploitation of H. thermocellum has been hampered due to the lack of simple and high-throughput genome engineering tools. Recently in our research group, a thermophilic bacterial CRISPR-Cas9-based system has been developed as a transcriptional suppression tool for regulation of gene expression. We applied ThermoCas9-based CRISPR interference (CRISPRi) to repress the H. thermocellum central metabolic lactate dehydrogenase (ldh) and phosphotransacetylase (pta) genes. The effects of repression on target genes were studied based on transcriptional expression and product formation. Single-guide RNA (sgRNA) under the control of native intergenic 16S/23S rRNA promoter from H. thermocellum directing the ThermodCas9 to the promoter region of both pta and ldh silencing transformants reduced expression up to 67% and 62% respectively. This resulted in 24% and 17% decrease in lactate and acetate production, correspondingly. Hence, the CRISPRi approach for H. thermocellum to downregulate metabolic genes can be used for remodelling of metabolic pathways without the requisite for genome engineering. These data established for the first time the feasibility of employing CRISPRi-mediated gene repression of metabolic genes in H. thermocellum DSM 1313.
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