Abstract
Fluorescence resonance energy transfer (FRET) is a popular tool to study intermoleculardistances and characterize structural or conformational changes of biological macromolecules.We investigate a novel inorganic/organic FRET pair with quantum dots (QDs) as donorsand DNA intercalating dyes, BOBO-3, as acceptors by using DNA as a linker.Typically, FRET efficiency increases with the number of stained DNA linked to aQD. However, with the use of intercalating dyes, we demonstrate that FRETefficiency at a fixed DNA:QD ratio can be further enhanced by increasing thenumber of dyes stained to a DNA strand through the use of an increased stainingdye/bp ratio. We exploit this flexibility in the staining ratio to maintain a high FRET efficiency of>0.90 despite a sixfold decrease in DNA concentration. Having characterized this newQD-mediated FRET system, we test this system in a cellular environment usingnanocomplexes generated by encapsulating DNA with commercial non-viral gene carriers.Using this novel FRET pair, we are able to monitor the configuration changes and fate ofthe DNA nanocomplexes during intracellular delivery, thereby providing an insight into themechanistic study of gene delivery.
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