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Tan, Ying-Xia; Chen, Chong; Wang, Ying-Li; Lin, Song; Wang, Yang; Li, Su-Bo; Jin, Xiao-Pan; Gao, Hong-Wei; Du, Fu-Sheng; Gong, Feng; Ji, Shou-Ping
The journal of gene medicine, 04/2012, Letnik: 14, Številka: 4Journal Article
Background Melittin is a commonly used cell‐penetrating peptide (CPP) for improving branched polyethylenimine (BPEI)‐mediated gene transfection. However, its application is limited owing to the cytotoxicity generated by the lytic activity at neutral pH. In the present study, we report two truncated peptides from melittin and florae with improved transfection efficiency. Methods Two truncated peptides consisting of 1–20 residues of melittin (MT20) and florae (FL20) were synthesized. Circular dichroism (CD) spectrometry was used to analyze the secondary structures of the peptides. The membrane‐lytic activity of the peptides and their potency in enhancing cellular uptake of calcein were evaluated. The peptides and BPEI mixtures were mixed with plasmid DNA to prepare peptide/BPEI/DNA complexes. The physicochemical characters of complexes were measured and the effect of the peptides on BPEI‐mediated transfection was determined. Results CD analysis and structure observation showed that the truncated peptides have α‐helical conformation, which was necessary for penetrating activity. The truncated peptides exhibited several advantages than their parent peptides: (i) they showed higher hemolytic potency in acidic pH but lower lytic activity than their parent peptides in neutral pH; (ii) enhanced calcein efficiently release from both early and late endosome; (iii) they did not affect the DNA‐binding affinity of BPEI and the physicochemical characteristics of BPEI/DNA complexes. Moreover, the peptides could increase BPEI‐mediated transfection efficiency in different cell lines (293FT, B16F10 and CHO‐K1) by simply mixing with BPEI, without causing cytotoxicity. Conclusions The results obtained in the present study indicate that the truncated peptides with higher endosomal disrupting activity were better enhancers for increasing transfection efficiency. Copyright © 2012 John Wiley & Sons, Ltd.
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