EFFICIENT METHOD OF PRION GENE SILENCING IN VITRO AND IN VIVO BY ANTISENSE-OLIGODEOXYNUCLEOTIDES CONJUGATED WITH NEW DIMETHYLAMINOETHYLMETHACRYLATE OLIGOELECTROLITE

L. A. Ivanytska, V. V. Stadnyk, Yu. V. Martyn, M. R. Kozak, S. V. Fedorova, O. V. Shtapenko, Yu. A. Rzhepetskyy, T. V. Skorokhoda, O. S. Zaichenko, V. V. Vlizlo


DOI: http://dx.doi.org/10.30970/sbi.0503.168

Abstract


The paper presents some issues of utilizing novel polymeric compound for impro­ving antisense-oligonucleotides delivery to mammalian cells. DNA-binding ability of dimethylaminoethylmethacrylate oligoelectrolite was tested. It was estimated that polyDMAEM in concentration less than 5 µg/ml did not induce cytotoxic effects in mouse embryonic fibroblasts and cells of L1210 line and could be used in in vitro studies. The study of polyDMAEM and its complexes with asODNs action on the cellular prion (PrPC) expression level evinced that different immobilized asODNs caused reduction of PrPC content by 70–90% in L1210 cells. It was found that polyDMAEM is able to decrease PrPC content by 40 % in L1210 cells by itself. Possible mechanisms of such influence are discussed. A possibility of successful using asODNs and carrier conjugates for cellular prion gene silencing in rat spleen and intestine is shown. Effects of polyDMAEM action on PrPC content has not been ascertained in vivo.


Keywords


antisense-oligodeoxynucleotides, gene silencing, prion, cationic oligo­electrolytes

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