Computational Design and Experimental Evaluation of MERS-CoV siRNAs in Selected Cell Lines.
Sayed Sartaj SohrabSherif Aly El-KafrawyZeenat MirzaAhmed M HassanFatima AlsaqafEsam Ibraheem AzharPublished in: Diagnostics (Basel, Switzerland) (2023)
Middle East respiratory syndrome coronavirus (MERS-CoV) is caused by a well-known coronavirus first identified in a hospitalized patient in the Kingdom of Saudi Arabia. MERS-CoV is a serious pathogen affecting both human and camel health globally, with camels being known carriers of viruses that spread to humans. In this work, MERS-CoV genomic sequences were retrieved and analyzed by multiple sequence alignment to design and predict siRNAs with online software. The siRNAs were designed from the orf1ab region of the virus genome because of its high sequence conservation and vital role in virus replication. The designed siRNAs were used for experimental evaluation in selected cell lines: Vero cells, HEK-293-T, and Huh-7. Virus inhibition was assessed according to the cycle threshold value during a quantitative real-time polymerase chain reaction. Out of 462 potential siRNAs, we filtered out 21 based on specific selection criteria without off-target effect. The selected siRNAs did not show any cellular toxicity in the tested cell lines at various concentrations. Based on our results, it was obvious that the combined use of siRNAs exhibited a reduction in MERS-CoV replication in the Vero, HEK-293-T, and Huh-7 cell lines, with the highest efficacy displayed in the Vero cells.
Keyphrases
- respiratory syndrome coronavirus
- sars cov
- coronavirus disease
- induced apoptosis
- saudi arabia
- cell cycle arrest
- endothelial cells
- oxidative stress
- public health
- health information
- signaling pathway
- case report
- mental health
- cell death
- magnetic resonance imaging
- gene expression
- risk assessment
- dna methylation
- candida albicans
- data analysis
- disease virus
- pi k akt