Yeast RNA (along with R848, Curdland, LPS and yeast cells in different conditions of culture) was used to induce DC activation.

This tumor RNA was used to transfect matured DC with the final aim to produce a DC vaccine.

Commercially available siRNA Smart Pool from Dharmacon targeted against the STAT3 transcription factor have been successfully transfected in human MDDCs.

Studies were carried out demonstrating that DC and other cell types can be activated in vitro by transfection with single stranded viral or synthetic RNA containing 5’ phosphates.

This shRNA was cloned into the pLVTHM lentiviral vector (obtained from the repository Addgene). This vector has been already successfully used to generate self-inactivating lentiviral vectors previously tested in DCs for transfection efficiency and absence of effects on their phenotype.

For transient transfection of synthetic siRNA, commercially available siRNA Smart Pool from Dharmacon targeted against the STAT3 transcription factor have been successfully transfected in human MDDCs (Lipofectamine 2000 as transfection reagent). The efficiency of silencing, as detected by western blot or FACS analysis of the protein was of 50-80% (depending on the donor), and peaked at 72 h post-transfection. Transfection efficiency was reproducibly higher than 80% and did not induce any type I IFN production.

There is siRNA available in the lab.

Electroporation of immature DCs with poly(I:C(12)U), a dsRNA analogue, resulted in phenotypic as well as functional changes, indicative of DC maturation.

The poly(I:C) was used for stimulating DCs. It was shown in mice that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC. Manufacturer: Invivogen Life Technologies

This is mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3). It was used to electroporate autologous dendritic cells in order to produce a vaccine for advanced melanoma patients.

The melanin-free total tumor RNA is obtained from melanoma metastases; it is suitable for in vitro amplification and DC transfection.

Co-electroporation of immature DCs with both poly(I:C(12)U) and Melan-A/MART-1 encoding mRNA induced strong anti-Melan-A/MART-1 CD8(+) T-cell responses in vitro. Higher numbers of Melan-A/MART-1-specific CTLs were consistently obtained with poly(I:C(12)U)-activated DCs compared to DCs matured in the presence of an inflammatory cytokine cocktail.

Total RNA was extracted from yeast cells after buffering, incubation and centrifugation by phenol extraction.

These constructs were used to transfect immature DCs.

This RNA was used to transfect matured DC with the final aim to produce a DC vaccine.

This construct was used as in the transfection of immature DC.