Vol 51(2017) N 5 p. 695-704; DOI 10.1134/S0026893317050065
B.P. Chelobanov1,2*, M.N. Repkova1,2, S.I. Baiborodin3, E.I. Ryabchikova1,2, D.A. Stetsenko1
Nuclear delivery of oligonucleotides via nanocomposites based on TiO2 nanoparticles and polylysine1Institute of Chemical Biology and Fundamental Medicine Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090 Russia
2Novosibirsk State University, Novosibirsk, 630090 Russia
3Institute of Cytology and GeneticsSiberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090 Russia
Received - 2016-07-05; Accepted - 2016-12-02
The nuclear delivery of nucleic acid derivatives is an essential prerequisite for successful antisense therapy. Using laser confocal and electron microscopy, we have studied the uptake of fluorescently labeled oligonucleotides in the form of nanocomposites with polylysine and TiO2 nanoparticles into Caco2, MDCK, and HeLa cells. In all three cell lines, bright fluorescence has been detected after 30 min in the nuclei (excluding the nucleoli) of the interphase cells; no substantial increase in the intensity of the signal was observed for next 24 hours. In all cells undergoing mitosis, the signal was localized in the cytoplasm with zones of higher intensity around chromatin. In some cells, at the beginning of interphase (G-1 phase), fluorescence was not detected at all. The latter may be explained by the brief moment in the cell cycle when oligonucleotides delivered in the nanocomposite cannot be taken up by cells. The studied nanocomposites are prone to aggregation. The degree of aggregation increases with the storage time up to complete loss of the ability of the nanocomposites to penetrate the cells.
deoxyribozyme (DNAzyme), cell delivery, scanning confocal laser microscopy, electron microscopy