Chimeric bifunctional oligonucleotides as a novel tool to invade telomerase assembly.
Bottom Line: Telomerase RNA and protein reverse transcriptase subunits are essential for the appearance of active telomerase in vitro.The approach is based on the application of chimeric bifunctional oligonucleotides that contain two oligonucleotide parts complementary to the functional domains of telomerase RNA connected with non-nucleotide linkers in different orientations (5'-3', 5'-5' or 3'-3').Such chimeras inhibited telomerase in vitro in the nM range, but were effective in vivo in sub-nM concentrations, predominantly due to their effect on telomerase assembly and dimerization.
Affiliation: Department of Chemistry and A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119992, Russian Federation Skolkovo Institute of Science and Technology, Novaya Street, 100, Skolkovo, Odintsovsky District, Moscow Region, 143025, Russian Federation.Show MeSH
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Mentions: M and J oligonucleotides alone showed a pronounced inhibitory effect on telomerase activity in vitro (IC50 100 nM and 19 nM, correspondingly) (Table 1). In case of M oligonucleotides, the position of c3 modification either at 5′- or 3′-end did not influence the inhibition (Figure 2A). N oligonucleotide did not show any detectable inhibitory effect in vitro. G oligonucleotide showed better inhibitory properties than corresponding M ones (IC50 11nM) (Table 1). For the 5′-3′ chimera Mc3N, the inhibitory effect was slightly better than for M alone (IC50 35 nM). However, when M and N were connected within a long oligonucleotide with c3 moved to the 3′-end (MNc3), no inhibitory effect was detected. In the case of the 5′-5′ chimera M*c3N (inverted M) and 3′-3′ chimera Mc3N* (inverted N) inhibitory activity increased approximately two times in comparison to Mc3N. For Nc3M chimera, inhibitory activity decreased (IC50 76 nM) (Figure 2A). In the case of J containing chimeras the inhibitory activity significantly (two times) reduced for Jc3N in comparison with J alone and was scarcely detectable in inverted forms. Mc3M showed very little inhibitory effect, although M*c3M (first M inverted) showed an IC50 value 11.9 nM significantly (7-fold) higher than M alone, and IC50 value of MMc3 was similar to the one for Mc3 (Figure 2A).
Affiliation: Department of Chemistry and A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119992, Russian Federation Skolkovo Institute of Science and Technology, Novaya Street, 100, Skolkovo, Odintsovsky District, Moscow Region, 143025, Russian Federation.