3 March 2009 Delivery of gene-expressing fragments using quantum dot
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Abstract
Gene therapy is an attractive approach to supplement a deficient gene function. Although there has been some success with specific gene delivery using various methods including viral vectors and liposomes, most of these methods have a limited efficiency or also carry a risk for oncogenesis. Fluorescent nanoparticles, such as nanocrystal quantum dots (QDs), have potential to be applied to molecular biology and bioimaging, since some nanocrystals emit higher and longer lasting fluorescence than conventional organic probes do. We herein report that quantum dots (QDs) conjugated with nuclear localizing signal peptides (NLSP) successfully introduced the gene-fragments with promoter elements, which promoted the expression of the enhanced green fluorescent protein (eGFP) gene in mammalian cells. The expression of eGFP protein was observed when the QD/geneconstruct was added to the culture media. The gene-expression efficiency varied depending on multiple factors around QDs, such as 1) the reading direction of gene fragments, 2) the quantity of gene fragments attached on the surface of QD-constructs, 3) the surface electronic charges varied according to the structure of QD/gene-constructs, and 4) the particle size of QD/gene complex varied according to the structure and amounts of gene fragments. Using this QD/geneconstruct system, eGFP protein could be detected 28 days after the gene-introduction whereas the fluorescence of QDs was disappeared. This system therefore provides another method for the intracellular delivery of gene-fragments without using either viral vectors or specific liposomes. These results suggest that inappropriate treatment and disposal of QDs may still have risks to the environmental pollution including human health under certain conditions. Here we propose the further research for the immune and physiological responses in not only immune cells but also other cells, in order to clear the effect of all other nanoscale products as well as nanocrystal QDs.
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Akiyoshi Hoshino, Akiyoshi Hoshino, Noriyoshi Manabe, Noriyoshi Manabe, Sanshiro Hanada, Sanshiro Hanada, Kouki Fujioka, Kouki Fujioka, Masato Yasuhara, Masato Yasuhara, Akihiko Kondo, Akihiko Kondo, Kenji Yamamoto, Kenji Yamamoto, } "Delivery of gene-expressing fragments using quantum dot", Proc. SPIE 7189, Colloidal Quantum Dots for Biomedical Applications IV, 71890V (3 March 2009); doi: 10.1117/12.808775; https://doi.org/10.1117/12.808775
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