TABLE 2.
siRNA specificitya
| Construct | Inhibition of:
|
|||
|---|---|---|---|---|
| Human DC-SIGN | Human L-SIGN | Chimpanzee DC-SIGN | Rhesus macaque DC-SIGN | |
| si-SIGN2 | + (0) | − (Ø) | + (0) | − (Ø) |
| si-SIGN3 | + (0) | + (1) | + (0) | + (0) |
| si-SIGN8 | + (0) | − (1) | − (1) | − (2) |
| si-SIGN11b | ++ (0/0/0/0/0/0) | ++ (2/2/2/2/2/2) | ++ (0/0/0/0/0/0) | ++ (1/0/0/2/2/5) |
| si-SIGN26 | − (0) | − (1) | − (1) | − (0) |
| si-SIGN35 | − (0) | − (1) | − (0) | − (1) |
Subconfluent 293T cells were cotransfected with pSUPER control plasmid, si-GFP, or si-SIGN constructs along with human DC-SIGN and pEGFP-N1 expression plasmids. Flow cytometric analysis of DC-SIGN expression was performed. The mean fluorescence intensity of DC-SIGN expression was measured in the GFP-positive cell population. To test for the specificity of si-SIGN constructs, the same assay was performed with the human DC-SIGN expression plasmid replaced by plasmids expressing L-SIGN or chimpanzee or rhesus macaque DC-SIGN. Percentages of inhibition relative to that under the pSUPER control experimental condition were then calculated. ++, strong inhibition; +, inhibition; −, no inhibition. The numbers within brackets correspond to mismatches between siRNAs and specific mRNA sequences. Ø, no significative homology.
si-SIGN11 targets six repeated sequences with 100% homology on the human DC-SIGN mRNA.