Silencing of Amyloid Precursor Protein Expression Using a New Engineered Delta Ribozyme
Figure 4
Relative APP mRNA level in HEK-293 cells expressing APP-SOFA-HDV Rz. The expression profile of APP in SOFA-HDV-Rz276-, -Rz753-, -Rz756-, and -Rz885-transfected HEK-293 cells. To assess APP knockdown efficiency at the mRNA level, quantitative real-time PCR was performed with each of the forward and reverse APP or GAPDH primers using the ABI Prism 7000 Sequence Detection System and SYBR Green DNA binding dye (Invitrogen). The specific amplification was assessed based on the dissociation curve profile. The APP gene expression profile was normalized against that of GAPDH. The quantitative PCR procedure was performed in duplicate in three independent reactions for each sample. **. An approximately 70–80% decrease in GAPDH-normalized APP mRNA levels was observed with cells expressing active SOFA-HDV Rz (the most potent Rz). The GAPDH-normalized levels of endogenous APP were not significantly altered in the untreated or empty vector-transfected control cells.
