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. 2023 Feb 5;31:494–511. doi: 10.1016/j.omtn.2023.02.005

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Rapid decrease in SpCas9 expression after transfection

(A) DNA repair efficiency blotted with percent SpCas9-positive cells quantified with immunostaining. Repair efficiency was predicted using ICE tool. All experiments were carried out in two independent repeats from patient 1. Statistical analysis was done using two-way ANOVA; p < 0.01. (B) Western blot for SpCas9 expression at days 2, 4, and 7 after transfection. TUBB was used as loading control. (C) Immunofluorescence staining of SpCas9 (red) at days 2, 4, and 7 after transfection. Nuclei were counterstained with Hoechst. Scale bars, 50 μm. All experiments were done in PHSats from patient 1.