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Updated: Jun 29, 2025

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
Combining a Base Deaminase Mutator with Phage-Assisted Evolution
Maryam Ali1, Raneem Akel1, Maria J Botero1
1Department of Chemistry, University of Toronto, Mississauga, ON, Canada.
Abstract:
Phage-assisted evolution has emerged as a powerful technique for improving a protein's function by using mutagenesis and selective pressure. However, mutations typically occur throughout the host's genome and are not limited to the gene-of-interest (GOI): these undesirable genomic mutations can yield host cells that circumvent the system's selective pressure. Our system targets mutations specifically toward the GOI by combining T7 targeted mutagenesis and phage-assisted evolution. This system improves the structure and function of proteins by accumulating favorable mutations that can change its binding affinity, specificity, and activity.
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