Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells
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RNA interference (RNAi) is a recently discovered phenomenon that employs the use of miRNA (microRNA) and siRNA (short interfering RNA) that can be utilized for gene therapy purposes. However, the problem that has arisen is delivery of the siRNAs into diseased cells. The compound chitosan has recently gained much attention because it possesses several properties that make it an ideal candidate as a delivery vehicle for siRNA. Chitosan/siRNA based nanoparticles have been synthesized and characterized that are less than 100 nm in diameter. Characterization of their size was completed by using transmission electron microscopy (TEM), scanning electron microscopy (SEM), and dynamic light scattering (DLS) techniques. Characterization of siRNA loading was accomplished by utilizing UV spectroscopy, EDS, and confocal microscopy to test encapsulation of siRNA. The colorectal cancer cell line HCT 116, was used to test transfection of siRNA across the cellular membrane. The transfection efficiencies were evaluated by using fluorescently tagged siRNA embedded within chitosan nanoparticles. The chitosan/siRNA delivery system was compared with the classical approach used for siRNA delivery; namely, siRNA in Lipofectamine 2000 as well as with cells that received no treatment. Live cell imaging was also employed to aid in visualization of transfection. The results show that the chitosan based systems do transfect siRNA across the cellular member.