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. 2020 Jan 13;21(2):502. doi: 10.3390/ijms21020502

Figure 5.

Figure 5

The dCas9-24XSunTag does not increase on-target DNA methylation. (A) Scheme of the dCas9-24XSunTag (dCS24) vector used in this study. (B) DNA methylation of individual CpG sites (on the x-axis) of the ISG15 region determined by targeted bis-seq after treatment of HEK293 cells with the dCS24, ISG15 sgRNA, and Ab-3A3L with the R831E, K844E, and R887E mutations in the DNMT3A part. Data from a single representative experiment are shown. (C) A mean methylation of the target (ISG15) and the off-target (VEGFA) regions analyzed by targeted bis-seq of HEK293 based on two independent experiments. Error bars show standard deviations. Note that higher methylation levels were observed with the dCas9-10XSunTag with the same Ab-3A3L mutants (Figure 3B). See also Figure S2.