Fig. 5. Co-delivery of Cas9 mRNA and sgRNA for in vitro gene editing in GFP⁺-HepG2 cells. RGD-lipid LNPs and C12-200 LNPs with various Cas9 mRNA/sgRNA ratios and LNP concentrations were selected. (a) GFP expression of HepG2 cells post treatment with LNPs co-delivering Cas9 mRNA and sgRNA. (b) GFP expression of HepG2 cells was assessed using flow cytometry. Cytometry plot shows decreased GFP expression in cells treated with 0.8 mg μL⁻¹ RGD-lipid LNPs and C12-200 LNPs at Cas9 mRNA/sgRNA of 4 : 1. (c) Quantification of flow cytometry shows that cells treated with RGD-lipid LNPs have lower GFP expression, compared to untreated cells, lipofectamine control, and C12-200 LNPs. ****p < 0.0001.