Fig. 3.

Fig. 3. Development and application of CRISPR-mediated microbial genome editing tools. (A) Major limitations of accurate CRISPR technologies: PAM-dependence, and mismatch tolerance. Red rectangles, and asterisks represent PAM sequences and mutated bases in the genome. (B) Development of single-base level CRISPR genome editing methods by target-mismatched sgRNAs and engineered Cas9 variants. Red asterisks represent single point mutations. (C) Application of microbial CRISPR genome editing in various fields: Basic biosciences (e.g., DNA metabolism in model microbes); Industrial biotechnology (e.g., engineering of microbial workhorses); Personalized human health (e.g., modification of gut microbiome); Agriculture and environment (e.g., regulation of plant-microbes interaction).

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