Synthetic mRNAs with Superior Translation and Stability Properties
The translational efficiency and stability of synthetic mRNA in both cultured cells and whole animals can be improved by incorporation of modified cap structures at the 5'-end. mRNAs are synthesized in vitro by a phage RNA polymerase transcribing a plasmid containing the mRNA sequence in the presence of all four NTPs plus a cap dinucleotide. Modifications in the cap dinucleotide at the 2'- or 3'-positions of m(7)Guo, or modifications in the polyphosphate chain, can improve both translational efficiency and stability of the mRNA, thereby increasing the amount and duration of protein expression. In the context of RNA-based immunotherapy, the latter is especially important for antigen producti…
The assessment of environmental pollution caused by mining and metallurgy wastes from highly polluted post-industrial regions in Southern Poland
Stored metallurgy and mining wastes contain relatively high amounts of potentially toxic elements. To monitor the distribution of contaminants originating from dumps, the chemical and physical properties of the wastes must be characterised. In this study, the chemical properties of wastes deposited in two different locations in Southern Poland (Szklary and Zloty Stok) were evaluated. Heaps located in Zloty Stok contain wastes from gold mineralisation comprising arsenic while wastes in Szklary originate from a factory that produced an iron-nickel alloy. In Szklary the total concentrations of Ca, Mg, Fe, Zn, Mn, Cr, Co, Cu, Ni, Tl, Ag, Cd and Pb were determined, while in Zloty Stok also As is…
Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo
Vaccination with in vitro transcribed RNA coding for tumor antigens is considered a promising approach for cancer immunotherapy and has already entered human clinical testing. One of the basic objectives for development of RNA as a drug is the optimization of immunobioavailability of the encoded antigen in vivo. By analyzing the effect of different synthetic 5' mRNA cap analogs on the kinetics of the encoded protein, we found that m(2)(7,2'-O)Gpp(S)pG (beta-S-ARCA) phosphorothioate caps, in particular the D1 diastereoisomer, profoundly enhance RNA stability and translational efficiency in immature but not mature dendritic cells. Moreover, in vivo delivery of the antigen as beta-S-ARCA(D1)-c…