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Intracellular disassembly and localization of a new P123-PEI-R13/DNA complex

Issue title: Frontiers in Biomedical Engineering and Biotechnology – Proceedings of the 3rd International Conference on Biomedical Engineering and Biotechnology, 25–28 September 2014, Beijing, China

Article type: Research Article

Authors: Zhu, Manman | Liu, Kehai; | Zhu, Qing | Chen, Shunsheng | Lv, Hui | Zhao, Wenfang | Mao, Yuan | Hu, Jing

Affiliations: Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, P.R. China

Note: [] Corresponding author: Kehai Liu, Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China. Tel.: +086-21-61900388; Fax: +086-21-61900365; E-mail: [email protected].

Keywords: P123-PEI-R13/DNA complex, disassembly, intracellular localization

DOI: 10.3233/BME-141001

Journal: Bio-Medical Materials and Engineering, vol. 24, no. 6, pp. 1925-1931, 2014

Published: 2014
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Abstract

The appropriate location and release of target gene is necessary for gene therapy. In our previous paper, a gene vector named P123-PEI-R13 has been successfully synthesized, and the physical characteristics and cellular trafficking of nanoparticle P123-PEI-R13/DNA has been explored explicitly, but little was known about its disassembly within cells. In order to investigate its intracellular disassembly, P123-PEI-R13/DNA complex was exposed to the different competitors (RNA, DNA, proteins) or different conditions of pH and osmolarity, DNA release was determined by gel electrophoresis. Meanwhile, confocal laser technology was used to locate the complex in cells. The results revealed that DNA, RNA and osmolarity could affect the stability of the complex obviously, especially RNA which exist in nucleus. In addition, the speed of DNA release decreased as the weight ratio of polymer increased. Images got by a confocal fluorescence microscope confirmed that after cell uptake, P123-PEI-R13 could translocate DNA into nucleus.

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