Teaching old dogs new tricks: genetic engineering methanogens

Methanogenic archaea, which are integral to global carbon and nitrogen cycling, currently face challenges in genetic manipulation due to unique physiology and limited genetic tools. This review provides a survey of current and past developments in the genetic engineering of methanogens, including se...

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Bibliographic Details
Published inApplied and environmental microbiology Vol. 90; no. 7; p. e0224723
Main Authors Myers, Tyler, Dykstra, Christy M
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 24.07.2024
Subjects
Archaea - genetics
Archaea - metabolism
CRISPR-Cas Systems
Gene Editing - methods
Genetic Engineering - methods
Genetics and Molecular Biology
Methane - metabolism
Minireview
Mutagenesis
genetic tools
CRISPR
Euryarchaeota
synthetic biology
transformation
mutagenesis
archaea
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Summary:Methanogenic archaea, which are integral to global carbon and nitrogen cycling, currently face challenges in genetic manipulation due to unique physiology and limited genetic tools. This review provides a survey of current and past developments in the genetic engineering of methanogens, including selection and counterselection markers, reporter systems, shuttle vectors, mutagenesis methods, markerless genetic exchange, and gene expression control. This review discusses genetic tools and emphasizes challenges tied to tool scarcity for specific methanogenic species. Mutagenesis techniques for methanogens, including physicochemical, transposon-mediated, liposome-mediated mutagenesis, and natural transformation, are outlined, along with achievements and challenges. Markerless genetic exchange strategies, such as homologous recombination and CRISPR/Cas-mediated genome editing, are also detailed. Finally, the review concludes by examining the control of gene expression in methanogens. The information presented underscores the urgent need for refined genetic tools in archaeal research. Despite historical challenges, recent advancements, notably CRISPR-based systems, hold promise for overcoming obstacles, with implications for global health, agriculture, climate change, and environmental engineering. This comprehensive review aims to bridge existing gaps in the literature, guiding future research in the expanding field of archaeal genetic engineering.
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The authors declare no conflict of interest.
ISSN:0099-2240
1098-5336
1098-5336
DOI:10.1128/aem.02247-23