Sohrab Sayed Sartaj, Aly El-Kafrawy Sherif, Mirza Zeenat, Hassan Ahmed M, Alsaqaf Fatima, Azhar Esam Ibraheem
Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Post Box No-80216, Jeddah 21589, Saudi Arabia.
Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
Saudi J Biol Sci. 2021 Feb;28(2):1348-1355. doi: 10.1016/j.sjbs.2020.11.066. Epub 2020 Nov 27.
The Middle East Respiratory Syndrome Coronavirus is well known to cause respiratory syndrome and this virus was identified and isolated for the first time from Jeddah, Saudi Arabia in 2012 from infected patient. In this report, we have conducted the prediction, designing and evaluation of siRNAs targeting Middle East Respiratory Syndrome Coronavirus orf1ab gene to inhibit the virus replication. By using bioinformatics software, total twenty-one functional, off-target reduced siRNA were selected from four hundred and sixty-two siRNAs based on their greater potency and specificity. We have evaluated only seven siRNAs to analyze their performance and efficacy as antivirals by reverse transfection approach in Vero cells. There was no cytotoxicity of siRNAs at various concentrations was observed in Vero cells. Based on the real-time PCR results, better inhibition of viral replication was observed in the siRNA-1 and 4 as compared to other siRNAs. The results generated from this work provided suitable information about the efficacy of siRNAs which encouraged us to further evaluate the remaining siRNAs to determine their inhibitory effect on the virus replication. We concluded that the insilico prediction and designing resulted in the screening of potential siRNAs with better efficiency, and strength. This can be used to develop oligonucleotide-based antiviral therapeutics against MERS-CoV in the near future.
中东呼吸综合征冠状病毒因可引发呼吸综合征而广为人知，该病毒于2012年首次在沙特阿拉伯吉达从一名感染患者体内被鉴定并分离出来。在本报告中，我们针对中东呼吸综合征冠状病毒的orf1ab基因进行了小干扰RNA（siRNA）的预测、设计及评估，以抑制病毒复制。通过使用生物信息学软件，基于其更高的效力和特异性，从462个siRNA中总共筛选出21个功能性、脱靶效应降低的siRNA。我们仅评估了7个siRNA，通过在Vero细胞中采用反向转染方法来分析它们作为抗病毒药物的性能和疗效。在Vero细胞中未观察到不同浓度的siRNA具有细胞毒性。基于实时荧光定量PCR结果，与其他siRNA相比，在siRNA - 1和4中观察到对病毒复制有更好的抑制作用。这项工作产生的结果提供了关于siRNA疗效的合适信息，这鼓励我们进一步评估其余的siRNA，以确定它们对病毒复制的抑制作用。我们得出结论，计算机模拟预测和设计导致筛选出了效率更高、效力更强的潜在siRNA。这可用于在不久的将来开发针对中东呼吸综合征冠状病毒的基于寡核苷酸的抗病毒疗法。
