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. Author manuscript; available in PMC: 2018 Jan 24.
Published in final edited form as: Nat Methods. 2017 Jul 24;14(9):891–896. doi: 10.1038/nmeth.4368

Figure 4. ciCas9 specificity and basal activity can be tuned.

Figure 4

(a) Editing is shown after 24 hours at the EMX1 on-target site (left panel) and an off-target site (right panel). ciCas9 and an enhanced-specificity variant (e-ciCas9) in the presence and absence of A3 are compared to wild type Cas9. Off-target editing with e-ciCas9 was not significantly increased relative to the no transfection control (one-sided t-test, n = 3, p = 0.21). Black bars depict means (n = 3 cell culture replicates). (b) Fluorescence polarization competitions between BH3 peptide variants and BODIPY-labeled BAK peptide for binding to BCL-xL. Data shown as inhibition of BODIPY-BAK binding. Error bars depict s.e.m. (n = 3 technical replicates). (c) Editing at the AAVS1 locus is shown for ciCas9 and two variants, L22 and F22, after 24 hours in the presence and absence of A3. Black bars depict means (n = 3 cell culture replicates).