Disruption of the HR6B enzyme expression and its effect on terminal erythroid differentiation
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The expression of Late Erythroblast-1 (LEB-1) increases during erythroid terminal differentiation (ETD). The hypothesis of this study was that the LEB-1 protein plays a role in the process of ETD. The aims of this study were (1) to optimize electroporation conditions and (2) to disrupt LEB-1 production using a siRNA expression cassette (SEC). Cultured Murine Erythroleukemic (MEL) Cells were used as a model of ETD. SEC's were prepared and transfected into MEL cells by electroporation. Control SEC's were used to differentiate non-specific effects of experimental manipulation on cell differentiation. A previously produced rabbit polyclonal antibody to LEB-1 was used to assess level of disruption of LEB-1 production in western blot assays. The results indicate the LEB-1 protein was not "knocked down" in induced MEL cells. However, the GAPDH positive control did not work indicating that the electroporation experiments will need to be further investigated.