Fig. 6. Intravenous delivery of LNP-Cas9/sgRNA by limb perfusion.

a Schematic illustration of mouse hindlimb and relevant muscle groups. The 3D model of hindlimb is adapted from ref. ⁵⁰. LNP-CRISPR was injected into the dorsal saphenous vein. b Effect of the injection volume (2.5–10 mL/kg) on exon skipping efficiency in each muscle group of the lower hindlimb following limb perfusion of LNP-CRISPR in hEx45-mdx44 mice. TA tibialis anterior, EDL extensor digitorum longus, GC gastrocnemius, PL plantaris, Sol soleus, PP planta pedis, QD quadriceps. Data are represented as means ± S.D. (n = 4 mice for limb perfusion injection groups, n = 3 mice for intramuscular injection group). p value by two-sided Dunnett’s test showed significantly different from intramuscular injection (2.5 mL/kg) in the same skeletal muscle. c Effect of the injection dose (1–10 mg/kg) of total RNA packaged in LNP under 5 mL/kg injection volume. Data are represented as means ± S.D. (n = 4 mice). p value by two-sided Williams’ test showed significantly different from limb perfusion-PBS of the same skeletal muscle. d Dose-dependent dystrophin restoration in TA and GC following 14 days of limb perfusion of LNP-CRISPR (n = 4 mice for each LNP injection group, n = 2 mice for PBS injection group).