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. 2018 Dec 18;13(12):e0209408. doi: 10.1371/journal.pone.0209408

Fig 3. Effect of linker length on methylation at the target site and at non-target sites.

Fig 3

(a) Frequency of methylation caused by dCas9-MC/MN with different linkers at the target CpG site at a gap length of 12 nucleotides from the PAM site. The P values are from Student’s t-tests (n = 4 for the 4-amino acid linker and n = 5 for the 15-amino acid linker). Four of the five measurements for the 15-amino acid linker are from [13] and one is new. (b) Reproducibility of the frequency of methylation at 482 C’s in 241 non-target CpG sites on plasmid pReporter. Data is for the 15-amino acid linker (repeat 1 is from our previous study [13]; repeat 2 is new). (c) Correlation of off-target methylation levels when using a 4-amino acid linker and 15-amino acid linker. (d) Correlation of off-target methylation levels when using a 15-amino acid linker and 25-amino acid linker. Frequencies were determined by high-throughput bisulfite sequencing.