ArticleMicrobial cell factories2019
Establishment and application of a CRISPR-Cas12a assisted genome-editing system in Zymomonas mobilis.
Article in Microbial cell factories, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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36 citing papers in PubMed, 89 citations in OpenAlex.
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- Genetic tools for engineering Zymomonas mobilis, Cereibacter sphaeroides and Novosphingobium aromaticivorans to improve production of bioenergy compounds.Microbial cell factories · 2025Review
- AI-powered high-throughput digital colony picker platform for sorting microbial strains by multi-modal phenotypes.Nature communications · 2025Article
- Precision targeting of genetic variations in mixed bacterial cultures using CRISPR-Cas12a-programmed λ phages.Frontiers in microbiology · 2025Article
- Paradigm of engineering recalcitrant non-model microorganism with dominant metabolic pathway as a biorefinery chassis.Nature communications · 2024Article
- Development of a starch-fermenting Zymomonas mobilis strain for bioethanol production.Microbial cell factories · 2024Article
- Comprehensive network of stress-induced responses in Zymomonas mobilis during bioethanol production: from physiological and molecular responses to the effects of system metabolic engineering.Microbial cell factories · 2024Review
- Biosensor-assisted CRISPRi high-throughput screening to identify genetic targets inSynthetic and systems biotechnology · 2024Article
- Metabolic engineering of an industrial bacteriumSynthetic and systems biotechnology · 2024Article
- Systematic investigation of TetR-family transcriptional regulators and their roles on lignocellulosic inhibitor acetate tolerance inFrontiers in bioengineering and biotechnology · 2024Article
- Microbial Cell Factories in the Bioeconomy Era: From Discovery to Creation.Biodesign research · 2024Review
- Characterization and Application of the Sugar Transporter Zmo0293 fromInternational journal of molecular sciences · 2023Article
- Article
- Metabolic engineering ofFrontiers in bioengineering and biotechnology · 2023Article
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
- Zymo-Parts: A Golden Gate Modular Cloning Toolbox for Heterologous Gene Expression inACS synthetic biology · 2022Article
- Identification and Characterization of Genes Related to Ampicillin Antibiotic Resistance inAntibiotics (Basel, Switzerland) · 2022Article
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13 authors at 1 institution in 1 country.
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Abstract
backgroundEfficient and convenient genome-editing toolkits can expedite genomic research and strain improvement for desirable phenotypes. Zymomonas mobilis is a highly efficient ethanol-producing bacterium with a small genome size and desirable industrial characteristics, which makes it a promising chassis for biorefinery and synthetic biology studies. While classical techniques for genetic manipulation are available for Z. mobilis, efficient genetic engineering toolkits enabling rapidly systematic and high-throughput genome editing in Z. mobilis are still lacking.
resultsUsing Cas12a (Cpf1) from Francisella novicida, a recombinant strain with inducible cas12a expression for genome editing was constructed in Z. mobilis ZM4, which can be used to mediate RNA-guided DNA cleavage at targeted genomic loci. gRNAs were then designed targeting the replicons of native plasmids of ZM4 with about 100% curing efficiency for three native plasmids. In addition, CRISPR-Cas12a recombineering was used to promote gene deletion and insertion in one step efficiently and precisely with efficiency up to 90%. Combined with single-stranded DNA (ssDNA), CRISPR-Cas12a system was also applied to introduce minor nucleotide modification precisely into the genome with high fidelity. Furthermore, the CRISPR-Cas12a system was employed to introduce a heterologous lactate dehydrogenase into Z. mobilis with a recombinant lactate-producing strain constructed.
conclusionsThis study applied CRISPR-Cas12a in Z. mobilis and established a genome editing tool for efficient and convenient genome engineering in Z. mobilis including plasmid curing, gene deletion and insertion, as well as nucleotide substitution, which can also be employed for metabolic engineering to help divert the carbon flux from ethanol production to other products such as lactate demonstrated in this work. The CRISPR-Cas12a system established in this study thus provides a versatile and powerful genome-editing tool in Z. mobilis for functional genomic research, strain improvement, as well as synthetic microbial chassis development for economic biochemical production.
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