Enhancement of transfection efficiency for HeLa cells via incorporating arginine moiety into chitosan

Dun Wan Zhu, Hai Ling Zhang, Jin Gen Bai, Wen Guang Liu, Xi Gang Leng, Cun Xian Song, Jian Yang, Xiao Wei Li, Xu Jin, Li Ping Song, Lan Xia Liu, Xiu Lan Li, Yang Zhang, Kang De Yao

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Arginine-rich peptides have attracted considerable attention due to their distinct internalization mechanism. It was reported that arginine and guanidino moieties were able to translocate through cell membranes and played a critical role in the process of membrane permeation. In this work, arginine was conjugated to the backbone of chitosan to form a novel chitosan derivative, arginine modified chitosan (Arg-CS). Arg-CS/DNA complexes were prepared according to the method of coacervation process. The physicochemical properties of Arg-CS and Arg-CS/DNA complexes were characterized and the transfection activity and efficiency mediated by Arg-CS/DNA complexes were investigated taking HeLa cells as target cells. Arg-CS was characterized by FTIR and 13C NMR. Arg-CS/DNA polyelectrolyte complexes were investigated by agarose gel retardation, dynamic light scattering (DLS) and atomic force microscopy (AFM). The results revealed that the Arg-CS/DNA complexes started to form at N/P ratio of 2:1, and the size of particles varied from 100 to 180 nm. The cytotoxicity of Arg-CS and their complexes with plasmid DNA were determined by MTT assay for HeLa cells, and the results suggested that Arg-CS/DNA complexes were slightly less toxic than Arg-CS. Moreover, the derivative alone and their complexes showed significantly lower toxicity than PEI and PEI/DNA complexes, respectively. Taking HeLa cells as target cells and using pGL3-control as reporter gene, the luciferase expression mediated by Arg-CS was greatly enhanced to about 100 folds compared with the luciferase expression mediated by chitosan at different pH media. These results suggest that Arg-CS is a promising candidate as a safe and efficient vector for gene delivery and transfection.

Original languageEnglish (US)
Pages (from-to)3207-3215
Number of pages9
JournalChinese Science Bulletin
Volume52
Issue number23
DOIs
StatePublished - Dec 1 2007

Keywords

  • Arginine-modified chitosan
  • Arginine-rich peptides
  • Chitosan
  • Gene delivery system
  • Non-viral vector

ASJC Scopus subject areas

  • General

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  • Cite this

    Zhu, D. W., Zhang, H. L., Bai, J. G., Liu, W. G., Leng, X. G., Song, C. X., Yang, J., Li, X. W., Jin, X., Song, L. P., Liu, L. X., Li, X. L., Zhang, Y., & Yao, K. D. (2007). Enhancement of transfection efficiency for HeLa cells via incorporating arginine moiety into chitosan. Chinese Science Bulletin, 52(23), 3207-3215. https://doi.org/10.1007/s11434-007-0455-y