Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia.
Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia.
Molecules. 2021 Apr 29;26(9):2610. doi: 10.3390/molecules26092610.
MERS-CoV was identified for the first time in Jeddah, Saudi Arabia in 2012 in a hospitalized patient. This virus subsequently spread to 27 countries with a total of 939 deaths and 2586 confirmed cases and now has become a serious concern globally. Camels are well known for the transmission of the virus to the human population. In this report, we have discussed the prediction, designing, and evaluation of potential siRNA targeting the ORF1ab gene for the inhibition of MERS-CoV replication. The online software, siDirect 2.0 was used to predict and design the siRNAs, their secondary structure and their target accessibility. ORF1ab gene folding was performed by RNAxs and RNAfold software. A total of twenty-one siRNAs were selected from 462 siRNAs according to their scoring and specificity. siRNAs were evaluated in vitro for their cytotoxicity and antiviral efficacy in Huh7 cell line. No significant cytotoxicity was observed for all siRNAs in Huh7 cells. The in vitro study showed the inhibition of viral replication by three siRNAs. The data generated in this study provide preliminary and encouraging information to evaluate the siRNAs separately as well as in combination against MERS-CoV replication in other cell lines. The prediction of siRNAs using online software resulted in the filtration and selection of potential siRNAs with high accuracy and strength. This computational approach resulted in three effective siRNAs that can be taken further to in vivo animal studies and can be used to develop safe and effective antiviral therapies for other prevalent disease-causing viruses.
MERS-CoV 于 2012 年在沙特阿拉伯吉达首次被发现，当时是一名住院患者。此后，该病毒传播到 27 个国家，共造成 939 人死亡，2586 人确诊，现已成为全球严重关切的问题。骆驼是众所周知的将病毒传播给人类的媒介。在本报告中，我们讨论了针对 ORF1ab 基因的潜在 siRNA 的预测、设计和评估，以抑制 MERS-CoV 的复制。在线软件 siDirect 2.0 用于预测和设计 siRNAs 及其二级结构和靶标可及性。ORF1ab 基因折叠由 RNAxs 和 RNAfold 软件进行。根据评分和特异性，从 462 个 siRNAs 中总共选择了 21 个 siRNAs。在 Huh7 细胞系中评估了 siRNAs 的细胞毒性和抗病毒功效。所有 siRNAs 在 Huh7 细胞中均未观察到明显的细胞毒性。体外研究表明，三种 siRNAs 抑制了病毒复制。本研究产生的数据提供了初步和令人鼓舞的信息，可分别评估 siRNAs 以及在其他细胞系中针对 MERS-CoV 复制的联合作用。在线软件对 siRNAs 的预测导致了高准确性和强度的潜在 siRNAs 的筛选和选择。这种计算方法产生了三种有效的 siRNAs，可以进一步进行体内动物研究，并可用于开发针对其他流行致病病毒的安全有效的抗病毒疗法。
