Re-examination of Peptide-Sequence-Dependent Gene Expression of Cysteine-Installed Pegylated Oligolysine/DNA
Yuichi Yamasaki1, Daiki Kumekawa1, Satoshi Yamauchi1
1Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8656, Japan.
Abstract:
We previously synthesized cysteine-installed C-terminally PEGylated oligolysines with 20 amino acid residues to form cross-linked polymeric micelles (PMs) with luciferase-coding plasmid DNA as a candidate for artificial gene vectors. Luciferase gene expression in HeLa cells mediated by PEG-CK18C, PEG-CK9CK9, and PEG-K9CK9C was reported to be 35-, 5.4-, and 1.3-fold higher than that mediated by cysteine-uninstalled PEGylated oligolysine PEG-K20, respectively. However, after the publication, the survival rate of HeLa cells used in the previous study was found to be lower than usual when subcutaneously implanted into mice to create a xenograft model. In this study, to re-examine the peptide sequence-dependent gene expression, gene expression efficacy mediated by PEG-peptide PMs was compared with the PM cellular uptake results using newly obtained HeLa cell lines and the additional cell lines Huh-7, PANC-1, and BxPC3. As a result, PEG-K9CK9C PMs mediated the maximum gene expression in all cell lines, and the corresponding cellular uptake was also obtained. Therefore, we concluded that our previous results were erroneously obtained due to normality-depleted HeLa cells. A comparison of physicochemical characterizations, gene expression efficacy, and cellular uptake of PEG-peptide PMs is discussed in detail.
