Labeling and intracellular tracking of functionally active plasmid DNA with semiconductor quantum dots

被引:94
作者
Srinivasan, Charudharshini
Lee, Jeunghoon
Papadimitrakopoulos, Fotios
Silbart, Lawrence K.
Zhao, Minhua
Burgess, Diane J.
机构
[1] Univ Connecticut, Dept Pharmaceut Sci, Storrs, CT 06269 USA
[2] Univ Connecticut, Inst Mat Sci, Dept Chem, Polymer Program,Nanomat Optoelect Lab, Storrs, CT 06269 USA
[3] Univ Connecticut, Ctr Excellence Vaccine Res, Dept Anim Sci, Storrs, CT 06269 USA
基金
美国国家科学基金会;
关键词
plasmid DNA; transfection; quantum dots; labeling; QD-DNA conjugates; DNA tracking; nuclear staining; gene expression;
D O I
10.1016/j.ymthe.2006.03.010
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Semiconductor nanocrystal quantum dots (QDs) allow long-term imaging in the cellular environment with high photostability. QD biolabeling techniques have previously been developed for tagging proteins and peptides as well as oligonucleotides. In this contribution, QD-decorated plasmid DNA was utilized for the first time for long-term intracellular and intranuclear tracking studies. Conjugation of plasmid DNA with phospholipid-coated QDs was accomplished using a peptide nucleic acid (PNA)-N-succinimidyl-3-(2-pyridylthio) propionate linker. Gel electrophoresis and confocal and atomic force microscopy (AFM) were used to confirm the structure of QD-DNA conjugates. AFM imaging also revealed that multiple QDs were attached in a cluster at the PNA-reactive site of the plasmid DNA. These QD-DNA conjugates were capable of expressing the reporter protein, enhanced green fluorescent protein, following transfection in Chinese hamster ovary (CHO-K1) cells with an efficiency of ca. 62%, which was comparable to the control (unconjugated) plasmid DNA.
引用
收藏
页码:192 / 201
页数:10
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