Article
Systematic in vitro profiling of off-target affinity, cleavage and efficiency for CRISPR enzymes.
Nucleic acids research - 21 May 2020
Zhang Liyang, Rube H Tomas, Vakulskas Christopher A, Behlke Mark A, Bussemaker Harmen J, Pufall Miles A
Abstract excerpt
CRISPR RNA-guided endonucleases (RGEs) cut or direct activities to specific genomic loci, yet each has off-target activities that are often unpredictable. We developed a pair of simple in vitro assays to systematically measure the DNA-binding specificity (Spec-seq), catalytic activity specificity (SEAM-seq) and cleavage efficiency of RGEs. By separately quantifying binding and cleavage specificity, Spec/SEAM-seq...
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