Fig. 4: Sustained anti-obese effect and improvement of pre-diabetic symptoms in DIO mice.

a Schematic of pharmacodynamic study treatments with saline control (Saline), scExe4 donor plasmid-loaded LNP only (Donor), scExe4 donor plasmid-loaded LNP plus Cas9 plasmid-loaded LNP (Donor+Cas9) and subcutaneous implantation of an osmotic pump filled with synthetic Exe4 peptide solution (Peptide) in the DIO mice model and mice fed with normal diet (Normal Diet). b Plasma Exe4 concentration profile in vivo scExe4 knock-in DIO mice. c Relative weekly food intake normalized to that of Saline. d Relative body weight normalized to that of Saline. e Glucose tolerance test (left panel) and insulin tolerance (right panel) result for DIO mice at 8, 16, and 24 weeks after dosing. f Plasma insulin levels (left panel) and HbA1c concentration (right panel) of DIO mice at 8, 16, and 24 weeks after dosing. c, d Asterisks (****p < 0.0001) denote a significant difference compared to Saline, Donor and Peptide using one-way ANOVA. No significant difference (no asterisk; p > 0.05) was observed between the Normal Diet and Donor+Cas9 groups at 24 weeks (d). e, f Asterisks (***p < 0.001, ****p < 0.0001) indicate significant differences for the Donor+Cas9 and Normal Diet group compared to other groups by one-way ANOVA. No significant difference (no asterisk; p > 0.05) was observed between the Donor+Cas9 and Normal Diet groups. Each line represents the group mean (n = 4 biological replicates), with each data point corresponding to an individual mouse.