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. 2019 Jun 12;5(6):eaaw8922. doi: 10.1126/sciadv.aaw8922

Fig. 6. Cytosolic delivery of Cas9 RNP for CRISPR-Cas9 gene editing.

Fig. 6

(A) P4-mediated Cas9/sgEGFP delivery for efficient EGFP genome editing. (B) DLS analysis of P4/Cas9 RNP nanoparticles. The inset is the TEM image of P4/Cas9 RNP nanoparticles. Scale bar, 500 nm. (C) Confocal images of 293T-EGFP cells treated with P4/RNP for 4 hours. Scale bar, 50 μm. (D) Flow cytometry analysis of 293T-EGFP cells treated with P4/RNP for 4 hours by means of mean fluorescence intensity. (E) Indel analysis from samples treated by P4/RNP. Commercial reagent CMAX was used as a positive control. T7E1 assay results from the intracellular delivery of P4/RNP targeting (F) AAVS1 and (G) HBB genes in 293T cells. (H) Genome editing efficiency, as revealed by the grayscale density of T7E1 results, after the intracellular delivery of RNP targeting EGFP locus (for 293T-EGFP cells), AAVS1 locus, and HBB locus (for 293T cells). Data represent mean ± SD (n = 3, Student’s t test, **P < 0.01). T7E1 assay results from the intracellular delivery of P4/RNP targeting (I) AAVS1 and (J) HBB genes in HCT-116 and HT-29 cells.