未经批准:吉达发现了一种新的冠状病毒,沙特阿拉伯于2012年被指定为中东呼吸综合征冠状病毒(MERS-CoV)。迄今为止,该病毒已在27个国家报告。已经报道了从骆驼向人类传播病毒。目前,目前尚无针对该病毒的疫苗或抗病毒疗法。
未经证实:siRNA是计算机预测的,设计,并以MERS-CoV-orf1ab区域为靶标化学合成。通过以两种不同浓度(10.0nM和5.0nM)在Vero细胞和HEK-293-T细胞系中递送作为转染试剂的Lipofectamine™2000和JetPRIMER的siRNA来实验评估抗病毒活性。定量实时PCR(qRT-PCR)的Ct值用于计算和确定从两种细胞系分离的细胞上清液和细胞裂解物中病毒RNA水平的降低。
UNASSIGNED:序列比对导致选择高度保守的区域。使用orf1ab区域来预测和设计siRNA,并且最终从通过在线软件产生的四百二十六个siRNA中选择总共二十一个siRNA。在两种浓度下,在两种细胞系中针对所选择的siRNA观察到病毒复制的抑制和病毒RNA的显著减少。根据Ct值,观察到siRNA#11、12、18和20在两种浓度的两种细胞系中表现最佳。
UNASSIGNED:根据结果和数据分析,结论是使用两种不同的转染试剂是显著有效的。但是发现Lipofectamine™2000是比JetPRIMER更好的转染试剂,用于在两种细胞系中递送siRNA。
UNASSIGNED: A new coronavirus was identified in Jeddah, Saudi Arabia in 2012 and designated as Middle East Respiratory Syndrome Coronavirus (MERS-CoV). To date, this virus has been reported in 27 countries. The virus transmission to humans has already been reported from camels. Currently, there is no vaccine or antiviral therapy available against this virus.
UNASSIGNED: The siRNAs were in silico predicted, designed, and chemically synthesized by using the MERS-CoV-orf1ab region as a target. The antiviral activity was experimentally evaluated by delivering the siRNAs with Lipofectamine™ 2000 and JetPRIMER as transfection reagents in both Vero cell and HEK-293-T cell lines at two different concentrations (10.0 nM and 5.0 nM). The Ct value of quantitative Real-Time PCR (qRT-PCR) was used to calculate and determine the reduction of viral RNA level in both cell supernatant and cell lysate isolated from both cell lines.
UNASSIGNED: The sequence alignment resulted in the selection of highly conserved regions. The orf1ab region was used to predict and design the siRNAs and a total of twenty-one siRNAs were finally selected from four hundred and twenty-six siRNAs generated by online software. Inhibition of viral replication and significant reduction of viral RNA was observed against selected siRNAs in both cell lines at both concentrations. Based on the Ct value, the siRNAs # 11, 12, 18, and 20 were observed to be the best performing in both cell lines at both concentrations.
UNASSIGNED: Based on the results and data analysis, it is concluded that the use of two different transfection reagents was significantly effective. But the Lipofectamine™ 2000 was found to be a better transfection reagent than the JetPRIMER for the delivery of siRNAs in both cell lines.