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. Author manuscript; available in PMC: 2016 Nov 1.
Published in final edited form as: Nat Protoc. 2016 Mar 31;11(5):853–871. doi: 10.1038/nprot.2016.043

Figure 5. Universal bait detection of off-targets for designed VEGFA gRNA.

Figure 5

(a) Circos plot (circos.ca) on custom log scale showing genome-wide profile of Cas9:SeC9-2 junctions in cycling v-Abl pro-B cells. Bin size is 5 Mb and 8751 unique junctions are shown from 2 independent libraries. Chromosomes are displayed as centromere to telomere in a clock-wise orientation. Blue lines link SeC9-2 off-targets to bait break-site while red lines link VEGFA off-targets to bait break-site. (b) List of identified off-targets for SeC9-2 or VEGFA. The off-targets were identified by MACS2 as described previously6. v-Abl cells (3×106) were nucleofected with a combined total of 3 μg plasmid DNA in SF solution using the DN-100 program (Lonza) and collected 48 hours post nucleofection.