Novel branched amphiphilic peptides for nucleic acids delivery

In this study, we synthesized branched amphiphilic peptides with different length of the branch sequences and distribution of hydrophobic and hydrophilic residues. Their applications in transfecting pDNA and mRNA were tested and we found that the branched peptides with longer hydrophobic branches and concentrated hydrophobic residues at the branch ends showed higher transfection efficiency and better than the commercial delivery vector PEI-25K at an optimal N/P ratio of 10 for pDNA delivery and N/P ratio of 16 for mRNA delivery. Highly efficient and safe non-viral vectors for nucleic acids delivery have attracted much attention due to their potential applications in gene therapy, gene editing and vaccination against infectious diseases, and various materials have been investigated and designed as delivery vectors. Herein, we designed a series of branched amphiphilic peptides (BAPs) and tested their applications as pDNA/mRNA delivery vectors. The BAP structure was inspired by the phospholipids, in which lysine oligomers were used as the “polar head”, segments containing phenylalanine, histidine and leucine were used as the “hydrophobic tails”, and a lysine residue was used as the branching point. By comparing the gel retardation, particle sizes and zeta potentials of the BAP/pDNA complexes of the short-branch BAPs (BAP-V1 ∼ BAP-V4), we determined the optimal lysine oligomer was K 6 . However, their cell transfection efficiencies were not satisfactory, and thus three long-branch BAPs (BAP-V5 ∼ BAP-V7) were further designed. In these long-branch BAPs, more hydrophobic residues were added and the overall amphiphilicity increased accordingly. The results showed that these three BAPs could effectively compact the nucleic acids, including both pDNA and mRNA, and all could transfect nucleic acids into HEK 293 cells, with low cytotoxicity. Among the three long-branch BAPs, BAP-V7 (bis(FFLFFHHH)-K-K 6 ) showed the best transfection efficiency at N/P = 10, which was better than the commercial transfection reagent PEI-25 K. These results indicate that increased amphiphilicity would also benefit for BAP mediated nucleic acid delivery. The designed BAPs provide more documents of such novel type of nucleic acids delivery vectors, which is worth of further investigation as a new gene theranostic platforms.