Figure 1a shows a significant decrease in the protein level of murine phospholipase D1 (mPLD1) in peripheral blood leukocytes of mice, following intravenous administration of siRNA (4 µg/mouse) to knockdown mPLD1 (Fig. 1A).
Studies of conditional knockout mice and/or siRNA treatment of either TANK1−/− or TANK2−/− cells could provide information on the nature of redundant function.
They report that, although an HDS-based injection of DNA complexed with PEI frequently causes the death of mice and decreases the expression of siRNA, the addition of pullulan to the DNA/PEI complex prevents the frequent deaths of the mice.
However, we could not correlate the sid-1 mRNA level with the uptake of siRNA into mouse tissues (data not shown), and it is thus likely that other transporters are also involved.
To test protection against WNV, peptide/siRNA complexes (at a peptide to siRNA molar ratio of 10∶1) were prepared in 100 200 µl of 5% glucose and injected intravenously at 50 µg of siRNA per mouse per injection 15 hours before i.p. infection with WNV or SLE (5LD50/mouse).
Although NG108-15 cells are mouse rat hybrid, transfection of siRNA for mouse KAP3 in PNG3 cells decreased the KAP3 protein level to ∼15% after 2 days (Supplementary Material, Fig. S3B).
5 × 10 CTLs were nucleofected with 3 μg of siRNA using Mouse T Nucleofector Kit (Amaxa) and X-001 program according to manufacturer's instructions.
This sensitive assay will allow studying further the pharmacokinetics and biodisponibility of siRNA injected in mice (Gilbert et al, in preparation).
In some experiments, the mice also received twice weekly injections of siRNA complexed with Oligofectamine into the tumor, or into the site of tumor implantation (prior to appearance of a visible tumor).
