In this study, polyplexes formed with our previously described lipo-xenopeptide carriers were evaluated for CRISPR/Cas9-based genome editing. Consisting of lipo-amino fatty acids (LAF) linked to succinoyl tetraethylene pentamine (Stp) containing backbone, these double pH-responsive carriers enabled co-complexation of Cas9 mRNA and sgRNA. Carriers of different topologies, LAF/Stp ratios and LAF types were evaluated. Up to 3-fold higher gene editing efficacies than gold standard Lipofectamine were reached in multiple different cell lines and reporter models, namely HepatdT:PCSK9, Hela GFPd2 and Hela mCherry-DMDEx23. Top performing U-shaped and bundle(B)-shaped carriers exhibited genome editing efficacies at sub-nanomolar EC₅₀ concentration of 0.4 nM sgRNA and 0.1 nM sgRNA, respectively, even after incubation in full (≥ 90 %) serum. Co-delivery of Cas9 mRNA, sgRNA and ssDNA resulted in over 35% BFP/GFP conversion in Hela GFPd2 by homology directed repair. Importantly, intravenous administration of polyplexes mediated in vivo editing of the dystrophin gene, triggering mRNA exon 23 splicing modulation in dystrophin-expressing cardiac muscle, skeletal muscle, and brain tissue in BALB/c mice.