Figure 4.
In vitro DNA cleavage by Cas9 nickases. (A) Schematic overview of the Cas9 nuclease and the paired Cas9 nickase. The PAM sequences and cleavage sites are shown in red. (B) Target sites in the human AAVS1 locus. The position of each target site is shown in red. (C) Schematic overview of DNA cleavage reactions. FAM dyes (shown in green) were linked to both 5′ ends of the DNA substrate. (D) DSBs and SSBs analyzed using fluorescent capillary electrophoresis. Fluorescently labeled DNA substrates were incubated with Cas9 nucleases or nickases before electrophoresis.
