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. 2016 Aug 30;6:32386. doi: 10.1038/srep32386

Figure 2. Off-target analysis of the multiplexed miRNA mutants.

Figure 2

Off-target gene sequences were identified as previously described19,24. Off-target sequences containing 1–2 nt mismatches (indicated by a lowercase letter highlighted in red) with each of the target sgRNA sequences are shown. Off-target activity was determined using a T7EI assay after PCR amplification of the off-target loci. miRNA on-target mutations were also amplified and processed with the T7EI assay as a positive read-out of the analysis. Black arrowheads indicate mutations in both on- and off-target PCR products with a frequency ≥10%. Mutation frequencies were quantified for each on-target and off-target sequence and plotted in the chart.