A highly efficient system enables rapid and precise C → T point mutations in cyanobacteria.
Cyanobacteria contribute to 25% of global carbon fixation through photosynthesis.
Conventional genetic manipulation methods are complicated and limit extensive research on cyanobacteria.
The developed base editing system allows for effective gene inactivation and precise point mutations.
Efficient multiplex editing is possible, expanding the capabilities for genetic modifications.
The editing system can be easily reversed using sucrose counter-selection.
Simplified
Cyanobacteria are important primary producers, contributing to 25% of the global carbon fixation through photosynthesis. They serve as model organisms to study the photosynthesis, and are important cell factories for synthetic biology. To enable efficient genetic dissection and metabolic engineering in cyanobacteria, effective and accurate genetic manipulation tools are required. However, genetic manipulation in cyanobacteria by the conventional homologous recombination-based method and the recently developed CRISPR-Cas gene editing system require complicated cloning steps, especially during multi-site editing and single base mutation. This restricts the extensive research on cyanobacteria and reduces its application potential. In this study, a highly efficient and convenient system was developed which allows rapid and precise C → T point mutation and gene inactivation in the genomes of Synechocystis and Anabaena. This base editing system also enables efficient multiplex editing and can be easily cured after editing by sucrose counter-selection. This work will expand the knowledge base regarding the engineering of cyanobacteria. The findings of this study will encourage the biotechnological applications of cyanobacteria.
Key numbers
90%
Editing Efficiency
Editing efficiency reached over 90% for specific genes after 20 days.
over 90% of genes
Targetable Genes
The pCyCBE system can inactivate over 90% of genes in cyanobacteria.
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