Fig. 5: Robust gene editing by Cas9/sgRNA RNP and meganuclease targeting PCSK9 using the VFIC and VEDIC systems. | Nature Communications

Fig. 5: Robust gene editing by Cas9/sgRNA RNP and meganuclease targeting PCSK9 using the VFIC and VEDIC systems.

From: Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors

Fig. 5

a Constructs generated for Cas9/sgRNA RNP delivery. b Schematic illustration regarding Cas9/sgRNA RNP encapsulation into engineered EVs. c Schematic of reporter cells used to functionally assess Cas9/sgRNA RNP delivery by engineered EVs. d Percentage of eGFP positive cells after addition of engineered EVs, as measured by flow cytometry 48, 72 and 96 h after EV addition. e Immunofluorescence demonstrated gene-editing in recipient cells after treatment with different doses of engineered EVs after 4 days. Scale bar, 100 µm, representative images. f Cas9 protein quantification in the engineered VEDIC and VFIC EVs. g Editing efficiency for the endogenous target mTTR in N2a cells by engineered EVs evaluated by Sanger sequencing. h Constructs generated for EV-mediated delivery of meganuclease targeting PCSK9. i WB analysis of PCSK9 and VSV-G protein in Huh7 cells after treatment with different doses of EVs in 24-well plates. Two-way ANOVA (Tukey) multiple comparisons test was used for analysis of (d). Experiments were done with 3 biological replicates and data are shown as mean ± SD. b Created in BioRender.com, Zheng, W. (2025) https://BioRender.com/g77t273. Exact statistical analysis was reported in the Source data and Source data are provided as a Source Data file.

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