TOP-SECRETS enables Cas9 nucleases to discriminate SNVs outside of PAMs

Abstract excerpt

<h4>ABSTRACT</h4> RNA-guided CRISPR nuclease Cas9 cannot reliably differentiate between single nucleotide variations (SNVs) of targeted DNA sequences determined by their guide RNA (gRNA): they typically exhibit similar nuclease activities at any of those variations, unless the variation occurs with specific sequence contexts known as protospacer adjacent motifs (PAMs). Our approach, “TOP-SECRETS,” generates gRNA...

Topics

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• CRISPR-Associated Protein 9
• CRISPR-Cas Systems
• Gene Editing
• Gene Targeting
• HEK293 Cells
• Mutagenesis
• Protein Domains
• Substrate Specificity

Identifiers and source

Literature Corpus work

ba415388-e050-532d-8943-5376cc1b8e10

DOI

10.1101/2025.05.06.652491