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Recycling gene carrier with high efficiency and low toxicity mediated by L-cystine-bridged bis(β-cyclodextrin)s.

Zhang YH, Chen Y, Zhang YM, Yang Y, Chen JT, Liu Y - Sci Rep (2014)

Bottom Line: The results showed that these nanometersized supramolecular crosslinking systems exhibited higher DNA transfection efficiencies and lower cytotoxicity than the commercial DNA carrier gold standard (25 kDa bPEI) for both normal cells and cancer cells, giving a very high DNA transfection efficiency up to 54% for 293T cells.Significantly, this type of supramolecular crosslinking system possesses a number of enzyme-responsive disulfide bonds, which can be cleaved by reductive enzyme to promote the DNA release but recovered by oxidative enzyme to make the carrier renewable.These results demonstrate that these supramolecular crosslinking systems can be used as promising gene carriers.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Tianjin 300071, P. R. China.

ABSTRACT
Constructing safe and effective gene delivery carriers is becoming highly desirable for gene therapy. Herein, a series of supramolecular crosslinking system were prepared through host-guest binding of adamantyl-modified low molecular weight of polyethyleneimine with L-cystine-bridged bis(β-cyclodextrin)s and characterized by (1)H NMR titration, electron microscopy, zeta potential, dynamic light-scattering, gel electrophoresis, flow cytometry and confocal fluorescence microscopy. The results showed that these nanometersized supramolecular crosslinking systems exhibited higher DNA transfection efficiencies and lower cytotoxicity than the commercial DNA carrier gold standard (25 kDa bPEI) for both normal cells and cancer cells, giving a very high DNA transfection efficiency up to 54% for 293T cells. Significantly, this type of supramolecular crosslinking system possesses a number of enzyme-responsive disulfide bonds, which can be cleaved by reductive enzyme to promote the DNA release but recovered by oxidative enzyme to make the carrier renewable. These results demonstrate that these supramolecular crosslinking systems can be used as promising gene carriers.

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Related in: MedlinePlus

Confocal fluorescence images of HeLa cells transfected with complexes for 24 h in the presence of serum.(a)–(c) cells were incubated with PEI-Ada-13.7@RDM-labled DNA; (d)–(f) cells were incubated with PEI-Ada-13.7-LCD@RDM-labled DNA. The plasmid DNA was labelled with rhodamine (RDM) and the nucleus were stained by DAPI (N/P = 20).
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f6: Confocal fluorescence images of HeLa cells transfected with complexes for 24 h in the presence of serum.(a)–(c) cells were incubated with PEI-Ada-13.7@RDM-labled DNA; (d)–(f) cells were incubated with PEI-Ada-13.7-LCD@RDM-labled DNA. The plasmid DNA was labelled with rhodamine (RDM) and the nucleus were stained by DAPI (N/P = 20).

Mentions: To gain more insight into the efficient cellular uptake of the supramolecular crosslinking system PEI-Ada-13.7-LCD, the fluorescent confocal images using rhodamine (RDM)-labeled pDNA (red) with HeLa cells as the model cell line was performed to study the intracellular localization, and DAPI (blue) was used to stain nucleus to observe the localization of carriers. As shown in Fig. 6, the cells treated by PEI-Ada-13.7-LCD exhibited stronger red fluorescence than those treated by PEI-Ada-13.7, indicating a higher DNA translocation efficiency of PEI-Ada-13.7-LCD. It should be noteworthy that the rhodamine-labled pDNA mainly located around the nucleus, which would be favourable to the transport of DNA through the karyotheca.


Recycling gene carrier with high efficiency and low toxicity mediated by L-cystine-bridged bis(β-cyclodextrin)s.

Zhang YH, Chen Y, Zhang YM, Yang Y, Chen JT, Liu Y - Sci Rep (2014)

Confocal fluorescence images of HeLa cells transfected with complexes for 24 h in the presence of serum.(a)–(c) cells were incubated with PEI-Ada-13.7@RDM-labled DNA; (d)–(f) cells were incubated with PEI-Ada-13.7-LCD@RDM-labled DNA. The plasmid DNA was labelled with rhodamine (RDM) and the nucleus were stained by DAPI (N/P = 20).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4265772&req=5

f6: Confocal fluorescence images of HeLa cells transfected with complexes for 24 h in the presence of serum.(a)–(c) cells were incubated with PEI-Ada-13.7@RDM-labled DNA; (d)–(f) cells were incubated with PEI-Ada-13.7-LCD@RDM-labled DNA. The plasmid DNA was labelled with rhodamine (RDM) and the nucleus were stained by DAPI (N/P = 20).
Mentions: To gain more insight into the efficient cellular uptake of the supramolecular crosslinking system PEI-Ada-13.7-LCD, the fluorescent confocal images using rhodamine (RDM)-labeled pDNA (red) with HeLa cells as the model cell line was performed to study the intracellular localization, and DAPI (blue) was used to stain nucleus to observe the localization of carriers. As shown in Fig. 6, the cells treated by PEI-Ada-13.7-LCD exhibited stronger red fluorescence than those treated by PEI-Ada-13.7, indicating a higher DNA translocation efficiency of PEI-Ada-13.7-LCD. It should be noteworthy that the rhodamine-labled pDNA mainly located around the nucleus, which would be favourable to the transport of DNA through the karyotheca.

Bottom Line: The results showed that these nanometersized supramolecular crosslinking systems exhibited higher DNA transfection efficiencies and lower cytotoxicity than the commercial DNA carrier gold standard (25 kDa bPEI) for both normal cells and cancer cells, giving a very high DNA transfection efficiency up to 54% for 293T cells.Significantly, this type of supramolecular crosslinking system possesses a number of enzyme-responsive disulfide bonds, which can be cleaved by reductive enzyme to promote the DNA release but recovered by oxidative enzyme to make the carrier renewable.These results demonstrate that these supramolecular crosslinking systems can be used as promising gene carriers.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Tianjin 300071, P. R. China.

ABSTRACT
Constructing safe and effective gene delivery carriers is becoming highly desirable for gene therapy. Herein, a series of supramolecular crosslinking system were prepared through host-guest binding of adamantyl-modified low molecular weight of polyethyleneimine with L-cystine-bridged bis(β-cyclodextrin)s and characterized by (1)H NMR titration, electron microscopy, zeta potential, dynamic light-scattering, gel electrophoresis, flow cytometry and confocal fluorescence microscopy. The results showed that these nanometersized supramolecular crosslinking systems exhibited higher DNA transfection efficiencies and lower cytotoxicity than the commercial DNA carrier gold standard (25 kDa bPEI) for both normal cells and cancer cells, giving a very high DNA transfection efficiency up to 54% for 293T cells. Significantly, this type of supramolecular crosslinking system possesses a number of enzyme-responsive disulfide bonds, which can be cleaved by reductive enzyme to promote the DNA release but recovered by oxidative enzyme to make the carrier renewable. These results demonstrate that these supramolecular crosslinking systems can be used as promising gene carriers.

Show MeSH
Related in: MedlinePlus