Assessment of the preventive effect of knockdown of cellular genes NXF1, PRPS1PRPS1 and NAA10 in influenza infection in an in vitro model.

Q3 Medicine
E A Pashkov, D A Shikvin, G A Pashkov, F G Nagieva, E A Bogdanova, A S Bykov, E P Pashkov, O A Svitich, V V Zverev
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引用次数: 0

Abstract

Introduction: Influenza is an acute respiratory viral infectious disease caused by the influenza viruses. Current preventive and therapeutic approaches are of great anti-epidemic importance, but there are a number of problems, such as the rapid emergence of resistant strains, the lack of cross-immunity and the effectiveness of vaccines. One of the approaches to the development of anti-influenza agents is the use of RNA interference and small interfering RNAs complementary to the mRNA target of viral and cellular genes. Aim ‒ to evaluate the prophylactic anti-influenza effect of siRNAs directed to the cellular genes NXF1, PRPS1 and NAA10 in an in vitro model.

Materials and methods: Antigenic variants of influenza A virus: A/California/7/09 (H1N1), A/WSN/33 (H1N1) and A/Brisbane/59/07 (H1N1); cell cultures A549 and MDCK. The study was performed using molecular genetic (transfection, NC isolation, RT-PCR-RV) and virological (cell culture infection, titration by visual CPE, viral titer assessment using the Ramakrishnan method) methods.

Results: It was shown that siRNAs targeting the cellular genes NXF1, PRPS1 and NAA10, when used prophylactically in cell culture at a concentration of 0.25 μg per well, during infection with influenza virus strains A/California/7/09 (H1N1), A/WSN/33 (H1N1) and A/Brisbane/59/07 (H1N1) at a multiplicity of infection of 0.01, reduced viral replication to a level of 220 TCID50 per 1 ml of cell medium, whereas in control untreated cells the viral yield was ~106 TCID50 per 1 ml of medium.

Conclusions: Reproduction of influenza A viruses directly depends on the protein products of the NXF1, PRPS1, and NAA10 genes. Reduced expression of these genes disrupts the life cycle and activity of influenza viruses. Such an approach can potentially be studied and used for closely and distantly related representatives of other virus families.

下调细胞基因NXF1、PRPS1PRPS1和NAA10对流感感染的预防作用
简介:流感是由流感病毒引起的急性呼吸道病毒性传染病。目前的预防和治疗方法对防治流行病具有重要意义,但存在许多问题,例如耐药菌株的迅速出现、缺乏交叉免疫和疫苗的有效性。开发抗流感药物的方法之一是使用RNA干扰和小干扰RNA来补充病毒和细胞基因的mRNA靶标。目的:在体外模型中评价细胞基因NXF1、PRPS1和NAA10的sirna的预防抗流感作用。材料与方法:甲型流感病毒抗原变异:A/California/7/09 (H1N1)、A/WSN/33 (H1N1)和A/Brisbane/59/07 (H1N1);细胞培养A549和MDCK。采用分子遗传学(转染、NC分离、RT-PCR-RV)和病毒学(细胞培养感染、视觉CPE滴定、Ramakrishnan法测定病毒滴度)方法进行研究。结果:结果表明,siRNAs针对细胞基因NXF1, PRPS1 NAA10,当在细胞培养用于预防在浓度为0.25μg /,在感染流感病毒株/加州/ 7/09 (H1N1), /网络/ 33 (H1N1)和布里斯班/ 59/07 (H1N1)感染复数的0.01,降低病毒复制水平的220 TCID50每毫升的细胞中,而在控制细胞治疗病毒产量~ 106 TCID50每毫升的媒介。结论:甲型流感病毒的繁殖直接依赖于NXF1、PRPS1和NAA10基因的蛋白产物。这些基因表达的减少会扰乱流感病毒的生命周期和活性。这种方法有可能被研究并用于其他病毒科的近亲和远亲代表。
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来源期刊
Voprosy virusologii
Voprosy virusologii Medicine-Infectious Diseases
CiteScore
2.00
自引率
0.00%
发文量
48
期刊介绍: The journal deals with advances in virology in Russia and abroad. It publishes papers dealing with investigations of viral diseases of man, animals and plants, the results of experimental research on different problems of general and special virology. The journal publishes materials are which promote introduction into practice of the achievements of the virological science in the eradication and incidence reduction of infectious diseases, as well as their diagnosis, treatment and prevention. The reader will find a description of new methods of investigation, new apparatus and devices.
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