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Effect of T4 DNA modifications of PAM-complementary cytosines on type II-A CRISPR–Cas sgRNA mediated DNA targeting. Cleavage assay of Cas9 on target DNA containing 5-hmC (indicated in red). Cas9 is loaded with either targeting sgRNA (T sgRNA) or non-targeting sgRNA (NT sgRNA). Restriction products of Cas9 are 57 and 33 bp. The marker is indicated by white arrows.
