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In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line

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 in-silico prediction, designing and evaluation of...

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Autores principales: Sohrab, Sayed Sartaj, Aly El-Kafrawy, Sherif, Mirza, Zeenat, Hassan, Ahmed M., Alsaqaf, Fatima, Azhar, Esam Ibraheem
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7833792/
https://www.ncbi.nlm.nih.gov/pubmed/33519276
http://dx.doi.org/10.1016/j.sjbs.2020.11.066
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author Sohrab, Sayed Sartaj
Aly El-Kafrawy, Sherif
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
author_facet Sohrab, Sayed Sartaj
Aly El-Kafrawy, Sherif
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
author_sort Sohrab, Sayed Sartaj
collection PubMed
description 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 in-silico 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.
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spelling pubmed-78337922021-01-26 In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line Sohrab, Sayed Sartaj Aly El-Kafrawy, Sherif Mirza, Zeenat Hassan, Ahmed M. Alsaqaf, Fatima Azhar, Esam Ibraheem Saudi J Biol Sci Original Article 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 in-silico 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. Elsevier 2021-02 2020-11-27 /pmc/articles/PMC7833792/ /pubmed/33519276 http://dx.doi.org/10.1016/j.sjbs.2020.11.066 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Sohrab, Sayed Sartaj
Aly El-Kafrawy, Sherif
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title_full In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title_fullStr In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title_full_unstemmed In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title_short In silico prediction and experimental validation of siRNAs targeting ORF1ab of MERS-CoV in Vero cell line
title_sort in silico prediction and experimental validation of sirnas targeting orf1ab of mers-cov in vero cell line
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7833792/
https://www.ncbi.nlm.nih.gov/pubmed/33519276
http://dx.doi.org/10.1016/j.sjbs.2020.11.066
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