ArticleMethods in enzymology2019
Kinetic characterization of Cas9 enzymes.
Article in Methods in enzymology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
What it found
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Who cites it
10 citing papers in PubMed.
- Biophysical characterization ofSensors & diagnostics · 2026Article
- Comparative characterization of Cas12f orthologs reveals mechanistic features underlying enhanced genome editing efficiency.Nature structural & molecular biology · 2026Article
- RepFluo, a Fast Fluorescent In Vitro Assay of Cas9 Activity Exploiting Melting Curve Analysis.ACS omega · 2025Article
- Rapid CRISPR-Cas9 target-strand nicking can provide phage resistance by reducing DNA abundance.Nucleic acids research · 2025Article
- DNA targeting by compact Cas9d and its resurrected ancestor.Nature communications · 2025Article
- Analysis of metal-dependent DNA nicking activities by Cas endonucleases.Methods in enzymology · 2025Article
- Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding.Plants (Basel, Switzerland) · 2023Review
- Microfluidic tool for rapid functional characterization of CRISPR complexes.New biotechnology · 2022Article
- Alignment of helicases on single-stranded DNA increases activity.Methods in enzymology · 2022Article
- A standard operating procedure for an enzymatic activity inhibition assay.European biophysics journal : EBJ · 2021Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Bacterial adaptive immune systems employ clustered regularly interspaced short palindromic repeats (CRISPR) along with their CRISPR-associated genes (Cas) to form CRISPR RNA (crRNA)-guided surveillance complexes, which target foreign nucleic acids for destruction. Cas9 is unique in that it is composed of a single polypeptide that utilizes both a crRNA and a trans-activating crRNA (tracrRNA) or a single guide RNA to create double-stranded breaks in sequences complementary to the RNA via the HNH and RuvC nuclease domains. Cas9 has become a revolutionary tool for gene-editing applications. Here, we describe methods for studying the cleavage activities of Cas9. We describe protocols for rapid quench-flow and stopped-flow kinetics and interpretation of the results. The protocols detailed here will be paramount for understanding the mechanistic basis for specificity of this enzyme, especially in efforts to improve accuracy for clinical use.
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Registered trials
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