Simultaneous Non-invasive Analysis of DNA Condensation and Stability by Two-step QD-FRET.
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Nanoscale vectors comprised of cationic polymers that condense DNA to form nanocomplexes are promising options for gene transfer. The rational design of more efficient nonviral gene carriers will be possible only with better mechanistic understanding of the critical rate-limiting steps, such as nanocomplex unpacking to release DNA and degradation by nucleases. We present a two-step quantum dot fluorescence resonance energy transfer (two-step QD-FRET) approach to simultaneously and non-invasively analyze DNA condensation and stability. Plasmid DNA, double-labeled with QD (525 nm emission) and nucleic acid dyes, were complexed with Cy5-labeled cationic gene carriers. The QD donor drives energy transfer stepwise through the intermediate nucleic acid dye to the final acceptor Cy5. At least three distinct states of DNA condensation and integrity were distinguished in single particle manner and within cells by quantitative ratiometric analysis of energy transfer efficiencies. This novel two-step QD-FRET method allows for more detailed assessment of the onset of DNA release and degradation simultaneously.
Published Version (Please cite this version)10.1016/j.nantod.2009.02.008
Publication InfoChen, Hunter H; Ho, Yi-Ping; Jiang, Xuan; Mao, Hai-Quan; Wang, Tza-Huei; & Leong, Kam W (2009). Simultaneous Non-invasive Analysis of DNA Condensation and Stability by Two-step QD-FRET. Nano Today, 4(2). pp. 125-134. 10.1016/j.nantod.2009.02.008. Retrieved from https://hdl.handle.net/10161/6974.
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Adjunct Professor of Biomedical Engineering
Professor Leong's research interest focuses on biomaterials design, particularly on synthesis of nanoparticles for DNA-based therapeutics, and nanostructured biomaterials for regenerative medicine Biomaterials Design: design of self-assembled fibers for tissue engineering microfluidics-mediated synthesis of multifunctional nanoparticles for drug and gene delivery synthesis of novel quantum dots for biomedical applications Con
This author no longer has a Scholars@Duke profile, so the information shown here reflects their Duke status at the time this item was deposited.