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.

The thermodynamic asymmetry of siRNA duplexes determines their silencing activity. Favorable asymmetry can be achieved by incorporation of mismatches into the 3' part of the sense strand, providing fork-siRNAs, which exhibit higher silencing activity and higher sensitivity to nucleases. Recently, we found that selective 2'-O-methyl modifications of the nuclease-sensitive sites of siRNA significantly improve its nuclease resistance without substantial loss of silencing activity. Here, we examined the impact of nucleotide mismatches and the number and location of 2'-O-methyl modifications on the silencing activity and nuclease resistance of anti-MDR1 siRNAs. We found that both nonmodified and selectively modified fork-siRNAs with 4 mismatches at the 3' end of the sense strand suppress the expression of target gene at lower effective concentrations than the parent siRNAs with classical duplex design. The selective modification of nuclease-sensitive sites significantly improved the stability of fork-siRNAs in the presence of serum. The selectively modified fork-siRNA duplexes provided inhibitory effect over a period of 12 days posttransfection, whereas the gene silencing activity of the nonmodified analogs expired within 6 days. Thus, selective chemical modifications and structural alteration of siRNA duplexes improve their silencing properties and significantly prolong the duration of their silencing effect.

Edited by Tamas DalmayKeywords: RNA interference siRNA Thermodynamic stability MDR1 2 0 -O-Methyl modification a b s t r a c tThe thermodynamic properties of siRNA duplexes are important for their silencing activity. siRNAs with high thermodynamic stability of both the central part of the duplex and in the whole, usually display low silencing activity. Destabilization of the central part of the siRNA duplex could increase its silencing activity. However, mismatches located in the central part of the duplex could substantially decrease the amount of RNAi efficacy, hindering active RISC formation and function. In this study, we examined the impact of duplex destabilization by nucleotide substitutions in the central part (7-10 nt counting from the 5 0 -end of the antisense strand) of the nuclease-resistant siRNA on its silencing activity.

The ongoing COVID-19 pandemic around the world and in Russia remains a major event of 2020. All over the world, research is being conducted to comprehensively study the patterns and manifestations of the epidemic  process. The main quantitative characteristics of SARS-CoV-2 transmission dynamics among the population, based on the data of official monitoring over the current situation, play an important role in the development of  the epidemiological surveillance system.The aim of this study is to explore the peculiarities of age-gender distribution of COVID-19 patients in Moscow.Material and methods. The data related to the epidemiological characteristics of age-gender structure of COVID-19 patients in Moscow between March 19, 2020 and April 15, 2020, at different stages of the  epidemic were retrospectively analyzed.Results and discussion. The mean age of COVID-19 patients in Moscow was 46,41±20,58 years. The gender ratio (male/female) among the patients was 52.7/47.3 %, wherein the indicators varied depending upon the  age. Male/female ratio in the age group “under 39” stood at 53.7/46.3 %, and “over 40 years of age” – at  39.3/60.7 %. The predominant age range among male cases was 19 to 39 years old – 35.4 %, while among female patients – 40–59 years (36.5 %). The age distribution of patients in Moscow is indicative of the fact that COVID-19 is a disease that primarily affects older age groups. The age structure of all COVID-19 cases during the observation period is characterized by predominance of adult patients over 19 years of age – 92,7 % (92,6–92,8 %), the share of patients aged 40–59 years is 35,7% (35,5–35,9 %). The differences in the age distribution in males and females are as follows: in the male cohort, the age groups 19–39 years old and 40–59 years old prevail – 35.4 % (35.1–35.7 %) and 34.9 % (34.6–35.2 %), respectively. The age group 40–59 years old – 36.5 % (36.3–36.8%) dominates in the female cohort.