Small-molecule inhibitors of proteasome increase CjCas9 protein stability
Fig 5
Bortezomib enhances the efficiency of CjCas9/ 2 sgRNA-mediated GAA repeat deletion in the FXN gene of HEK293T cells.
A) Schematic representation of the human FXN gene with the positions of the sgRNAs used in our experiment. B) Schematic description of the experiment: cells were co-treated with or without bortezomib (4 nM) and a plasmid encoding CjCas9 under CMV promoter and 2 sgRNAs under U6 promoters. C) The number of plasmid copies was determined by qPCR. D) The level of CjCas9 protein was quantified by western blot and normalized with β-actin. E) 72 hours after transfection, gene editing efficiency on FXN gene was measured by ddPCR. Data are means ± SEM (n ≥ 3), *p < 0.05, **p < 0.005, and ***p < 0.0005 (Student’s t tests).