Silencing activity of 2′-O-methyl modified anti-MDR1 siRNAs with mismatches in the central part of the duplexes

Petrova, Natalya S.; Meschaninova, Mariya I.; Venyaminova, Alya G.; Zenkova, Marina A.; Vlassov, Valentin V.; Chernolovskaya, E. L.

doi:10.1016/j.febslet.2011.06.003

Cited by 14 publications

(10 citation statements)

References 35 publications

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“…These are degradation of siRNA by serum nucleases, binding of siRNA with serum protein, siRNA accumulation in non-target organs and tissues, excretion by the kidneys, low bioavailability, and inefficient silencing within the cells 11 . To study the performance of anti- MDR1 siRNA and its conjugate with cholesterol in vivo, we chose siRNA targeted to the 411–431 nt region of human MDR1 mRNA (siMDR), shown in our previous study to have the highest bioactivity in cell culture 25 . 2′-O-methyl modifications were introduced into nuclease-sensitive sites according to the algorithm developed by us previously 23 .…”

Section: Resultsmentioning

confidence: 99%

Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene

Chernikov

Gladkikh

Meschaninova

et al. 2017

Molecular Therapy - Nucleic Acids

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Chemical modifications are an effective way to improve the therapeutic properties of small interfering RNAs (siRNAs), making them more resistant to degradation in serum and ensuring their delivery to target cells and tissues. Here, we studied the carrier-free biodistribution and biological activity of a nuclease-resistant anti-MDR1 cholesterol-siRNA conjugate in healthy and tumor-bearing severe combined immune deficiency (SCID) mice. The attachment of cholesterol to siRNA provided its efficient accumulation in the liver and in tumors, and reduced its retention in the kidneys after intravenous and intraperitoneal injection. The major part of cholesterol-siRNA after intramuscular and subcutaneous injections remained in the injection place. Confocal microscopy data demonstrated that cholesterol-siRNA spread deep in the tissue and was present in the cytoplasm of almost all the liver and tumor cells. The reduction of P-glycoprotein level in human KB-8-5 xenograft overexpressing the MDR1 gene by 60% was observed at days 5–6 after injection. Then, its initial level recovered by the eighth day. The data showed that, regardless of the mode of administration (intravenous, intraperitoneal, or peritumoral), cholesterol-siMDR efficiently reduced the P-glycoprotein level in tumors. The designed anti-MDR1 conjugate has potential as an adjuvant therapeutic for the reversal of multiple drug resistance of cancer cells.

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Section: Resultsmentioning

confidence: 99%

Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene

Chernikov

Gladkikh

Meschaninova

et al. 2017

Molecular Therapy - Nucleic Acids

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“…In 2010 Maier and co-workers reported that the modulation of the thermal stability can enhance the potency of siRNA molecules [39]. The destabilization of the siRNA duplex could be achieved by the introduction of chemical modifications or on account of base pair mismatches [40,41]. The melting data stressed the destabilizing effects of the T L presence on the siRNA duplex (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Modulation of the RNA Interference Activity Using Central Mismatched siRNAs and Acyclic Threoninol Nucleic Acids (aTNA) Units

2015

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Abstract:The understanding of the mechanisms behind nucleotide recognition by Argonaute 2, core protein of the RNA-induced silencing complex, is a key aspect in the optimization of small interfering RNAs (siRNAs) activity. To date, great efforts have been focused on the modification of certain regions of siRNA, such as the 3'/5'-termini and the seed region. Only a few reports have described the roles of central positions flanking the cleavage site during the silence process. In this study, we investigate the potential correlations between the thermodynamic and silencing properties of siRNA molecules carrying, at internal positions, an acyclic L-threoninol nucleic acid (aTNA) modification. Depending on position, the silencing is weakened or impaired. Furthermore, we evaluate the contribution of mismatches facing either a natural nucleotide or an aTNA modification to the siRNA potency. The position 11 of the antisense strand is more permissive to mismatches and aTNA modification, in respect to the position 10. Additionally, comparing the ON-/OFF-target silencing of central mismatched siRNAs with 5'-terminal modified siRNA, we concluded: (i) central perturbation of duplex pairing features weights more on potency rather than silencing asymmetry; (ii) complete bias for the ON-target silencing can be achieved with single L-threoninol modification near the 5'-end of the sense strand. OPEN ACCESSMolecules 2015, 20 7603

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“…4 and 6 days post‐transfection, the culture medium was removed and the cells were lysed in 60 μl of Sample buffer (Sigma–Aldrich, USA). Western blotting was performed as described with [12] (detailed information in Supplementary material).…”

Section: Methodsmentioning

confidence: 99%

42‐ and 63‐bp anti‐MDR1‐siRNAs bearing 2′‐OMe modifications in nuclease‐sensitive sites induce specific and potent gene silencing

et al. 2014

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Edited by Tamas DalmayKeywords: small interfering RNA Dicer-substrate RNAs Interferon response 2 0 -O-methyl modification a b s t r a c t DsRNAs longer than 30 bp induce interferon response and global changes in gene expression profile in mammalians. 21 bp siRNA and 25/27 bp dsiRNA acting via RNA interference mechanism are used for specific gene silencing in this class of organisms. We designed selectively 2 0 -O-methyl-modified 42 and 63 bp anti-MDR1-siRNAs that silence the expression of P-glycoprotein and restore the sensitivity of drug-resistant cancer cells to cytostatic more efficiently than canonical 21 bp siRNAs. We also show that they act in a Dicer-independent mode and are devoid of immunostimulating properties. Our findings suggest that 42 and 63 bp siRNAs could be used as potential therapeutics.

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Silencing activity of 2′-O-methyl modified anti-MDR1 siRNAs with mismatches in the central part of the duplexes

Cited by 14 publications

References 35 publications

Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene

Cholesterol-Containing Nuclease-Resistant siRNA Accumulates in Tumors in a Carrier-free Mode and Silences MDR1 Gene

Modulation of the RNA Interference Activity Using Central Mismatched siRNAs and Acyclic Threoninol Nucleic Acids (aTNA) Units

42‐ and 63‐bp anti‐MDR1‐siRNAs bearing 2′‐OMe modifications in nuclease‐sensitive sites induce specific and potent gene silencing

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