Simultaneous Non-invasive Analysis of DNA Condensation and Stability by Two-step QD-FRET.

Loading...
Thumbnail Image

Date

2009-04-01

Journal Title

Journal ISSN

Volume Title

Repository Usage Stats

252
views
555
downloads

Citation Stats

Abstract

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.

Department

Description

Provenance

Subjects

Citation

Published Version (Please cite this version)

10.1016/j.nantod.2009.02.008

Publication Info

Chen, Hunter H, Yi-Ping Ho, Xuan Jiang, Hai-Quan Mao, Tza-Huei Wang and Kam W Leong (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.

This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.


Unless otherwise indicated, scholarly articles published by Duke faculty members are made available here with a CC-BY-NC (Creative Commons Attribution Non-Commercial) license, as enabled by the Duke Open Access Policy. If you wish to use the materials in ways not already permitted under CC-BY-NC, please consult the copyright owner. Other materials are made available here through the author’s grant of a non-exclusive license to make their work openly accessible.