ABSTRACT
We report the development of dendritic siRNA nanostructures that are able to penetrate even difficult to transfect cells such as neurons with the help of a special receptor ligand. The nanoparticles elicit strong siRNA responses, despite the dendritic structure. An siRNA dendrimer directed against the crucial rabies virus (RABV) nucleoprotein (Nâ protein) and phosphoprotein (Pâ protein) allowed the suppression of the virus titer in neurons below the detection limit. The cell-penetrating siRNA dendrimers, which were assembled using click chemistry, open up new avenues toward finding novel molecules able to cure this deadly disease.
Subject(s)
Dendrimers , Nanostructures , RNA, Small Interfering/chemistry , Chromatography, High Pressure Liquid , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Click chemistry of alkyne-modified RNA with different receptor ligand azides was used to prepare 3'-folate, 3'-cholesterol, and, as a new entity, 3'-anandamide-modified RNA in high yields and excellent purity. The anandamide-modified RNA shows surprisingly high transfection properties and enables the delivery of siRNA even into difficult-to-transfect RBL-2H3 cells which model neuronal uptake. Furthermore, the system was employed in human immune cells (BJAB), demonstrating silencing effects similar to those of a cationic, benchmark transfection reagent. In addition, the anandamide conjugates were found to be nontoxic. The reported chemistry and the described properties of the anandamide siRNA extend the possibilities of using siRNA-based gene silencing in neuronal and immune cells.
Subject(s)
Arachidonic Acids/chemistry , B-Lymphocytes/metabolism , Endocannabinoids/chemistry , Gene Silencing , Neurons/metabolism , Polyunsaturated Alkamides/chemistry , RNA, Small Interfering/administration & dosage , RNA, Small Interfering/chemistry , Animals , Base Sequence , Cell Line , Cholesterol/chemistry , Click Chemistry , Folic Acid/chemistry , HeLa Cells , Humans , Luciferases/genetics , RNA, Small Interfering/genetics , Renilla/enzymologyABSTRACT
Stable cationic carotenoid aggregates - predominantly of the J-type - develop when the hydrochlorides of carotenoid aldoximes and ketoximes are exposed to water. The oxime hydrochlorides are obtained by simple syntheses from commercially available food color carotenoids. Bluish-purple, unstable transient compounds were observed during hydrochlorination performed at liquid nitrogen temperature.
