Bacterial directed evolution of CRISPR base editors.

Reilly Q Mach; Shannon M Miller

doi:10.1016/bs.mie.2025.01.003

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Bacterial directed evolution of CRISPR base editors.

Reilly Q Mach·Shannon M Miller

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Abstract

Base editing and other precision editing agents have transformed the utility and therapeutic potential of CRISPR-based genome editing. While some native enzymes edit efficiently with their nature-derived function, many enzymes require rational engineering or directed evolution to enhance the compatibility with mammalian cell genome editing. While many methods of engineering and directed evolution exist, plate-based discrete evolution offers an ideal balance between ease of use and engineering power. Here, we describe a detailed method for the bacterial directed evolution of CRISPR base editors that compounds technical ease with flexibility of application.

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Gene Editing Directed Molecular Evolution CRISPR-Cas Systems Escherichia coli Clustered Regularly Interspaced Short Palindromic Repeats Base editing CRISPR Directed evolution Precision genome editing Protein engineering Journal Article

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Reilly Q Mach, Shannon M Miller. 2025-03-08. Bacterial directed evolution of CRISPR base editors.. https://doi.org/10.1016/bs.mie.2025.01.003

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