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. 2016 Feb 16;24(3):475–487. doi: 10.1038/mt.2016.1

Figure 5.

Figure 5

Strategies to reduce off-target events. (a) Modification of the FokI domain to prevent homodimerization of ZFN or TALEN monomers. (b) Modification of the Cas9 nuclease to generate a nicking version of Cas9 (Cas9N). Cas9N can generate single-stranded DNA breaks. (c) Inactivation of the Cas9 endonuclease to create a dead Cas9. Fusion of the FokI domain creates a dCas9-FokI enzyme that requires a pair of dCas9-FokI to achieve dimerization of the FokI domain for DNA cleavage. (d) Cas9 orthologs with longer protospacer adjacent motif sequences can result in less potential off-target sites in the genome.