ArticleFrontiers in cell and developmental biology2021
A Highly Sensitive GFP Activation Assay for Detection of DNA Cleavage in Cells.
Article in Frontiers in cell and developmental biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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3 citing papers in PubMed.
- Enhanced genome editing with a Streptococcus equinus Cas9.Communications biology · 2025Article
- Engineering of a high-fidelity Cas12a nuclease variant capable of allele-specific editing.PLoS biology · 2024Article
- Phage-assisted evolution of compact Cas9 variants targeting a simple NNG PAM.Nature chemical biology · 2024Article
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7 authors.
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Abstract
CRISPR/Cas9 nucleases hold great potential for gene therapy, but they frequently induce unwanted off-target cleavage. We previously developed a GFP activation assay for detection of DNA cleavage in cells. Here, we demonstrate two novel applications of this assay. First, we use this assay to confirm off-target cleavage that cannot be detected by targeted deep sequencing in cells before. Second, we use this approach to detect multiple alternative PAMs recognized by SpCas9. These noncanonical PAMs are associated with low cleavage activity, but targets associated with these PAMs must be considered as potential off-target sites. Taken together, the GFP activation assay is a powerful platform for DNA cleavage detection in cells.
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