2020/08/24 by Ahmed E. Dhamad, Daniel J. Lessner · 36 citations
Biochemistry, Genetics and Molecular Biology · Environmental Science · #Archaea #Bacteria #Biology #CRISPR and Genetic Engineering #Computational biology #Euryarchaeota #Gene #Genetics #Methanogen #Methanogenesis #Methanosarcina #Microbial Fuel Cells and Bioremediation #Microbial Metabolic Engineering and Bioproduction #Nitrogen fixation #Nitrogenase
paper · open access · doi:10.1128/aem.01402-20
published in Applied and Environmental Microbiology 86(21) (American Society for Microbiology)
openalex publication_date 2020/08/24 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/06
Genetic tools are needed to understand and manipulate the biology of archaea, which serve critical roles in the biosphere. Methanogenic archaea (methanogens) are essential for the biological production of methane, an intermediate in the global carbon cycle, an important greenhouse gas, and a biofuel. The CRISPRi-dCas9 system in the model methanogen Methanosarcina acetivorans is, to our knowledge, the first Cas9-based CRISPR interference system in archaea. Results demonstrate that the system is remarkably efficient in targeted gene repression and provide new insight into nitrogen fixation by methanogens, the only archaea with nitrogenase. Overall, the CRISPRi-dCas9 system provides a simple, yet powerful, genetic tool to control the expression of target genes and operons in methanogens.