Small Interfering RNA-Mediated Gene Silencing in T Lymphocytes
Journal of Immunology
FIGURE 3. A restrictive number of designed siRNA sequences are effective at silencing. A, Effect of three different siRNAs targeting CD8␣. Effective concentrations of three different CD8␣ siRNAs were evaluated by titrating increasing amounts of the siRNAs into the cuvettes before electroporation. CD8␣ expression was evaluated 36 h posttransfection, and is plotted as a percentage of wild-type levels. Effective siRNA (ࡗ); two ineffective siRNAs (OE, f). B, Relative target locations of the siRNAs
... ions of the siRNAs used in these studies. Schematic of CD4 and CD8␣ mRNAs (not to scale), showing ORF and 5Ј-and 3Ј-UTR. ϩ, Indicates siRNAs that are effective. FIGURE 4. Effective and ineffective siRNAs can compete for silencing. Effective CD8␣ (A) and CD4 (B) siRNA-mediated silencing is competed by increasing concentration of cotransfected ineffective CD4 and CD8␣ siRNAs. Effective CD8␣ (C) or CD4 (D) siRNA-mediated silencing is competed by increasing concentrations of cotransfected ineffective CD8␣ or CD4 siRNA. Red indicates E10 cells transfected with 2.5 mol effective siRNA and black is the nontransfected control. Green and blue indicate the addition of half the amount of an ineffective siRNA.