Biochemical and Biophysical Research Communications
Cationic liposome-mediated delivery of siRNAs in adult mice
Section snippets
Materials and methods
Reagents. 1,2 Dioleoyl-3-trimethylammonium-propane (DOTAP) was purchased from Roche Diagnostics. FITC-labeled siRNAs were chemically synthesized and purified by Eurogentec. The siRNAs were annealed in transfection buffer (20 mM Hepes, 150 mM NaCl, pH 7.4) at 3 μg/μl.
Preparation of siRNA–liposome complexes for i.v. injection of mice. For each mice 20–100 μg of siRNAs in 60 μl of transfection buffer was transferred into a sterile Eppendorf tube. In a separate sterile polystyrene tube 50 μg of DOTAP was
Cationic liposome-mediated intravenous nucleic acid delivery
Similar to antisense oligonucleotides and ribozymes, synthetic siRNAs are generally delivered to cells via liposome-based transfection reagents [6]. These reagents offer the possibility of developing pharmaceutical compounds for local or systemic delivery. Notably, most patients who die from cancer have subclinical metastatic disease present at the time of diagnosis [9]. Thus, it is essential that molecular medicine-based therapies to treat cancer be adjusted to a systemic administration. In
Discussion
The study of nucleic acids has revealed remarkable properties of RNA molecules that could make them attractive therapeutic agents, independent of their well-known ability to encode biologically active proteins [13]. This application is now mainly driven by small interfering RNAs that serve as guide sequences to induce target-specific mRNA cleavage via the activation of a highly conserved regulatory mechanism, called RNA interference or posttranscriptional gene silencing [3]. However, crucial
Acknowledgements
This work was supported by the Norwegian Cancer Society. We thank Dr. Anne Dybwad for critical reading of the manuscript and Khu Ky Cuong for excellent technical assistance.
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