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Bacterial directed evolution of CRISPR base editors
Reilly Q Mach1, Shannon M Miller1
1The Scripps Research Institute.
Methods in Enzymology
|March 22, 2025
Summary
Directed evolution enhances CRISPR base editors for mammalian genome editing. This method uses bacterial evolution for efficient and flexible base editor engineering.
Area of Science:
- Molecular Biology
- Biotechnology
- Genome Engineering
Background:
- CRISPR-based genome editing, including base editing, has advanced therapeutic potential.
- Many CRISPR enzymes require engineering for efficient mammalian genome editing.
Purpose of the Study:
- To describe a method for bacterial directed evolution of CRISPR base editors.
- To enhance the compatibility of base editors with mammalian cells.
Main Methods:
- Plate-based discrete evolution was employed for enzyme engineering.
- Bacterial systems were utilized for directed evolution of CRISPR base editors.
Main Results:
- The described method offers a balance of ease of use and engineering power.
- The bacterial directed evolution approach provides flexibility for base editor applications.
Conclusions:
- Bacterial directed evolution is an effective strategy for engineering CRISPR base editors.
- This method facilitates the development of more compatible and potent genome editing tools.
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