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Title: A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots

Abstract

Highlights: ► An easy and direct way to prepare QDs–DNA complexes was developed. ► Surface charge of QDs was tuned with different ratio of amino and glycolate. ► Transfection efficiency was dependent on the surface zeta potentials of QDs. ► Cellular toxicity of this gene vectors is much lower than commercial liposome. ► Whole intracellular behavior of QDs–DNA complexes can be monitored in real time. -- Abstract: Nanoparticle carrier has been developed by combining water-soluble quantum dots and plasmid DNA expressed enhanced green fluorescent protein (EGFP) in a convenient and direct way. First the QDs with different surface charges were obtained by coating with amino and carboxyl terminals at different ratios. Then plasmid DNA was conjugated to QDs via electrostatic interaction. The resultant QDs–DNA complexes showed enhanced resistance to DNase I digestion. The following transfection experiments demonstrated that the transfection efficiency was dependent on the surface charges on QDs. The real time imaging of the transfection process showed that the nanoparticles experienced binding, penetrating the cell membrane and entering cytoplasm in the first 6 h of transfection. The green fluorescence of EGFP began to appear after 18 h transfection and plasmid DNA was fully expressed in the following 6 h.more » This new QDs–DNA platform showed great potential as new gene delivery carrier.« less

Authors:
; ;  [1];  [2];  [3];  [1];  [2];  [3];  [1];  [2]
  1. Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074 (China)
  2. (China)
  3. Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074 (China)
Publication Date:
OSTI Identifier:
22215570
Resource Type:
Journal Article
Journal Name:
Materials Research Bulletin
Additional Journal Information:
Journal Volume: 47; Journal Issue: 11; Other Information: Copyright (c) 2012 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0025-5408
Country of Publication:
United States
Language:
English
Subject:
77 NANOSCIENCE AND NANOTECHNOLOGY; CELL MEMBRANES; CYTOPLASM; DNA; FLUORESCENCE; PHOTOELECTRON SPECTROSCOPY; QUANTUM DOTS; SURFACES

Citation Formats

Zhang, Hai-Li, Zhang, Ming-Zhen, Li, Xiang-Yong, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Wan, Min, Li, Yong-Qiang, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Zhang, Rong-Ying, Zhao, Yuan-Di, E-mail: zydi@mail.hust.edu.cn, and Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074. A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots. United States: N. p., 2012. Web. doi:10.1016/J.MATERRESBULL.2012.07.030.
Zhang, Hai-Li, Zhang, Ming-Zhen, Li, Xiang-Yong, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Wan, Min, Li, Yong-Qiang, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Zhang, Rong-Ying, Zhao, Yuan-Di, E-mail: zydi@mail.hust.edu.cn, & Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074. A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots. United States. doi:10.1016/J.MATERRESBULL.2012.07.030.
Zhang, Hai-Li, Zhang, Ming-Zhen, Li, Xiang-Yong, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Wan, Min, Li, Yong-Qiang, Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074, Zhang, Rong-Ying, Zhao, Yuan-Di, E-mail: zydi@mail.hust.edu.cn, and Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074. Thu . "A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots". United States. doi:10.1016/J.MATERRESBULL.2012.07.030.
@article{osti_22215570,
title = {A convenient method of preparing gene vector for real time monitoring transfection process based on the quantum dots},
author = {Zhang, Hai-Li and Zhang, Ming-Zhen and Li, Xiang-Yong and Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074 and Wan, Min and Li, Yong-Qiang and Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074 and Zhang, Rong-Ying and Zhao, Yuan-Di, E-mail: zydi@mail.hust.edu.cn and Key Laboratory of Biomedical Photonics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Department of Biomedical Engineering, Wuhan 430074},
abstractNote = {Highlights: ► An easy and direct way to prepare QDs–DNA complexes was developed. ► Surface charge of QDs was tuned with different ratio of amino and glycolate. ► Transfection efficiency was dependent on the surface zeta potentials of QDs. ► Cellular toxicity of this gene vectors is much lower than commercial liposome. ► Whole intracellular behavior of QDs–DNA complexes can be monitored in real time. -- Abstract: Nanoparticle carrier has been developed by combining water-soluble quantum dots and plasmid DNA expressed enhanced green fluorescent protein (EGFP) in a convenient and direct way. First the QDs with different surface charges were obtained by coating with amino and carboxyl terminals at different ratios. Then plasmid DNA was conjugated to QDs via electrostatic interaction. The resultant QDs–DNA complexes showed enhanced resistance to DNase I digestion. The following transfection experiments demonstrated that the transfection efficiency was dependent on the surface charges on QDs. The real time imaging of the transfection process showed that the nanoparticles experienced binding, penetrating the cell membrane and entering cytoplasm in the first 6 h of transfection. The green fluorescence of EGFP began to appear after 18 h transfection and plasmid DNA was fully expressed in the following 6 h. This new QDs–DNA platform showed great potential as new gene delivery carrier.},
doi = {10.1016/J.MATERRESBULL.2012.07.030},
journal = {Materials Research Bulletin},
issn = {0025-5408},
number = 11,
volume = 47,
place = {United States},
year = {2012},
month = {11}
}