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. 2019 Sep 24;28(13):3510–3522.e5. doi: 10.1016/j.celrep.2019.08.065

Figure 2.

Figure 2

Off-Target Analysis in Mouse HSPCs

(A) On-target and 5 highest-risk off-target sites of sgLmnb1 and sgActb as predicted by CrispRGold.

(B) T7EI assays show editing activity of sgLmnb1 (left) and sgActb (right) in the on- and/or off-target sites from control or treated HSPCs (ND, not determined).

(C) ICE analysis showing the indel frequency in the on- and/or off-targeted sites. Data are shown as means ± SD from three independent experiments.

(D and E) Activated-Sca1+ HSPCs from C57BL/6 mice were electroporated with RNPs containing the indicated sgRNAs and infected with the indicated donor viruses. The frequency of mCherry+ (D) and BFP+ (E) LSK cells was determined by flow cytometry based on three independent experiments. Data are shown as means ± SD (∗∗p < 0.01, Mann-Whitney test).