Cargando…

In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells

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 vi...

Descripción completa

Detalles Bibliográficos
Autores principales: El-Kafrawy, Sherif Aly, Sohrab, Sayed Sartaj, Mirza, Zeenat, Hassan, Ahmed M., Alsaqaf, Fatima, Azhar, Esam Ibraheem
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125306/
https://www.ncbi.nlm.nih.gov/pubmed/33947034
http://dx.doi.org/10.3390/molecules26092610
_version_ 1783693462460170240
author El-Kafrawy, Sherif Aly
Sohrab, Sayed Sartaj
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
author_facet El-Kafrawy, Sherif Aly
Sohrab, Sayed Sartaj
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
author_sort El-Kafrawy, Sherif Aly
collection PubMed
description 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.
format Online
Article
Text
id pubmed-8125306
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-81253062021-05-17 In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells El-Kafrawy, Sherif Aly Sohrab, Sayed Sartaj Mirza, Zeenat Hassan, Ahmed M. Alsaqaf, Fatima Azhar, Esam Ibraheem Molecules Article 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. MDPI 2021-04-29 /pmc/articles/PMC8125306/ /pubmed/33947034 http://dx.doi.org/10.3390/molecules26092610 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
El-Kafrawy, Sherif Aly
Sohrab, Sayed Sartaj
Mirza, Zeenat
Hassan, Ahmed M.
Alsaqaf, Fatima
Azhar, Esam Ibraheem
In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title_full In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title_fullStr In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title_full_unstemmed In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title_short In Vitro Inhibitory Analysis of Rationally Designed siRNAs against MERS-CoV Replication in Huh7 Cells
title_sort in vitro inhibitory analysis of rationally designed sirnas against mers-cov replication in huh7 cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125306/
https://www.ncbi.nlm.nih.gov/pubmed/33947034
http://dx.doi.org/10.3390/molecules26092610
work_keys_str_mv AT elkafrawysherifaly invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells
AT sohrabsayedsartaj invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells
AT mirzazeenat invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells
AT hassanahmedm invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells
AT alsaqaffatima invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells
AT azharesamibraheem invitroinhibitoryanalysisofrationallydesignedsirnasagainstmerscovreplicationinhuh7cells