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Oxazole Yellow Homodimer YOYO-1-labeled DNA: A Fluorescent Complex That Can Be Used to Assess Structural Changes in DNA Following Formation and Cellular Delivery of Cationic Lipid DNA Complexes

Oxazole Yellow Homodimer YOYO-1-labeled DNA: A Fluorescent Complex That Can Be Used to Assess Structural Changes in DNA Following Formation and Cellular Delivery of Cationic Lipid DNA Complexes

M Wong, S Kong, W H Dragowska, M B Bally

Biochim Biophys Acta. 2001 Jul 2;1527(1-2):61-72.

PMID: 11420144

Abstract:

To improve transfection efficiency following delivery of plasmid expression vectors using lipid-based carriers, it is crucial to define structural characteristics of the lipid/DNA complexes that optimize transgene expression. Due to its strong affinity for DNA and high quantum yield, the fluorescent DNA intercalator YOYO-1 was used as a tool to assess changes in DNA that occur following lipid binding and cell delivery. In this study, the stability of the dye/DNA complex following binding of poly-L-lysine or monocationic lipids is characterized. More than 98% of the fluorescence measured for a defined DNA/YOYO-1 complex was lost when DNA was condensed using poly-L-lysine. This loss in fluorescence could be attributed to displacement of bound dye. In contrast, more than 30% of the fluorescence of the dye-labeled DNA was retained after formation of cationic lipid/DNA complexes. Significantly, the results illustrate differences in structural changes cationic lipids and PLL exert on plasmid DNA. The fluorescent lipid/DNA complex was used to assess DNA delivery to murine B16/BL6 cells in vitro. An assay relying on fluorescence resonance energy transfer between bound YOYO-1 and propidium iodide was used to distinguish between DNA attached to the cell surface and internalized DNA.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP152068092	Oxazole yellow		152068-09-2	Price

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