Spacer Extension Reconciles Specificity and Activity in High-Fidelity Cas9 Genome Editing

Abstract excerpt

<h4> Abstract </h4> Engineering high-fidelity CRISPR enzymes often leads to reduced cleavage activity, creating a significant hurdle in balancing nuclease specificity and efficiency for clinical applications. Here, we demonstrate that extending the spacer to 21 or 22 nucleotides restores the impaired cleavage activity of SuperFi-Cas9, an engineered high-fidelity Cas9 variant with seven mutations at the PAM-dista...

Topics

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• CRISPR and Genetic Engineering
• CRISPR-Associated Protein 9
• CRISPR-Cas Systems
• Cryoelectron Microscopy
• Endonucleases
• Models, Molecular
• Mutation
• RNA, Guide, CRISPR-Cas Systems

Identifiers and source

Literature Corpus work

b3696128-7e02-54b8-a79f-49e5bf2feea7

DOI

10.1101/2025.01.16.633458