Improving prime editor function through targeted protein changes in yeast

Delivery of the optimized prime editing variant PE_Y18 increases editing rates up to 3.5-fold compared to PEmax.

• OrthoRep, a yeast-based platform, was used to enhance the editing efficiency of prime editors.
• Multiple mutations that positively affect prime editing activity were identified after several rounds of evolution.
• The most effective mutations were combined to create the variant PE_Y18.
• PE_Y18 showed improved editing rates when delivered into mammalian cell lines and in vivo into the liver or brain.
• The study provides proof-of-concept for optimizing genome editing tools using directed protein evolution.

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