Cholesterol-modified anti-MDR1 small interfering RNA: Uptake and biological activity

Kruglova, Natal’ya S.; Meschaninova, Mariya I.; Ven’yaminova, A. G.; Zenkova, Marina A.; Vlassov, Valentin V.; Chernolovskaya, E. L.

doi:10.1134/s002689331002010x

Cited by 7 publications

(5 citation statements)

References 34 publications

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“…Exposure to siRNAs caused a concentration-dependent reduction of the P-glycoprotein level. The concentration-dependent results are consistent with our previous observations using anti-MDR1 siR-NAs, which can effectively inhibit target gene expression at a concentration of 200 nM [24]. To compare the biological activity of siRNAs with different lengths we used concentrations of the equal ''dose'' of the 21 nucleotide sequence of siMDR-M repeated two or three times (dose-equivalent concentrations), presuming the processing of siMDR-D and siMDR-T into two or three molecules of siMDR-M, respectively.…”

Section: Discussionsupporting

confidence: 92%

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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Section: Discussionsupporting

confidence: 92%

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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“…The pronounced reduction of the number of living cells was observed at Day 6 of cells incubation with cholesterol-containing siMDR ( Figure 6 ). This effect was similar to the effect observed at Day 4 after cell transfection with siMDR/Lipophectamine 2000 ( 54 ).…”

Section: Discussionsupporting

confidence: 83%

“…These modifications improved the bio-distribution of siRNAs ( 26 , 37 ) and significantly increased their cellular uptake ( 25 , 27 , 35 ). The presence of the cholesterol in siRNA did not however guarantee efficient cellular uptake and gene silencing without the aid of the transfection agent ( 54 ). Moreover, the data on the efficiency of carrier-free uptake and the biological effects of lipophilic siRNAs vary significantly in different reports where different synthetic methods for the introduction of lipophilic moieties resulted in the obtaining of the conjugates with different structures ( 26 , 37 , 54 ).…”

Section: Discussionmentioning

confidence: 99%

Carrier-free cellular uptake and the gene-silencing activity of the lipophilic siRNAs is strongly affected by the length of the linker between siRNA and lipophilic group

Petrova¹,

Chernikov²,

Meschaninova

et al. 2011

Nucleic Acids Research

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The conjugation of siRNA to molecules, which can be internalized into the cell via natural transport mechanisms, can result in the enhancement of siRNA cellular uptake. Herein, the carrier-free cellular uptake of nuclease-resistant anti-MDR1 siRNA equipped with lipophilic residues (cholesterol, lithocholic acid, oleyl alcohol and litocholic acid oleylamide) attached to the 5′-end of the sense strand via oligomethylene linker of various length was investigated. A convenient combination of H-phosphonate and phosphoramidite methods was developed for the synthesis of 5′-lipophilic conjugates of siRNAs. It was found that lipophilic siRNA are able to effectively penetrate into HEK293, HepG2 and KB-8-5 cancer cells when used in a micromolar concentration range. The efficiency of the uptake is dependent upon the type of lipophilic moiety, the length of the linker between the moiety and the siRNA and cell type. Among all the conjugates tested, the cholesterol-conjugated siRNAs with linkers containing from 6 to 10 carbon atoms demonstrate the optimal uptake and gene silencing properties: the shortening of the linker reduces the efficiency of the cellular uptake of siRNA conjugates, whereas the lengthening of the linker facilitates the uptake but retards the gene silencing effect and decreases the efficiency of the silencing.

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“…Conjugates of anti-mdr1 siRNA sense strand [36,37] with trileucine ( 19 ) and cholesteryl-6-aminohexylcarbamate ( 20 ) were also successfully synthesized in optimal conditions (Table 1, Scheme 3A). Moreover, the proposed approach allows, when necessary, to lengthen the spacer between the oligonucleotide and the ligand by several successive activation/addition solid-phase reactions through DSC.…”

Section: Resultsmentioning

confidence: 99%

Novel Convenient Approach to the Solid-Phase Synthesis of Oligonucleotide Conjugates

et al. 2019

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A novel and convenient approach for the solid-phase 5′-functionalization of oligonucleotides is proposed in this article. The approach is based on the activation of free 5′-hydroxyl of polymer support-bound protected oligonucleotides by N,N′-disuccinimidyl carbonate followed by interaction with amino-containing ligands. Novel amino-containing derivatives of closo-dodecaborate, estrone, cholesterol, and α-tocopherol were specially prepared. A wide range of oligonucleotide conjugates bearing closo-dodecaborate, short peptide, pyrene, lipophilic residues (cholesterol, α-tocopherol, folate, estrone), aliphatic diamines, and propargylamine were synthesized and characterized to demonstrate the versatility of the approach. The developed method is suitable for the conjugate synthesis of oligonucleotides of different types (ribo-, deoxyribo-, 2′-O-methylribo-, and others).

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Cholesterol-modified anti-MDR1 small interfering RNA: Uptake and biological activity

Cited by 7 publications

References 34 publications

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

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

Carrier-free cellular uptake and the gene-silencing activity of the lipophilic siRNAs is strongly affected by the length of the linker between siRNA and lipophilic group

Novel Convenient Approach to the Solid-Phase Synthesis of Oligonucleotide Conjugates

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