Version 1
: Received: 27 February 2019 / Approved: 28 February 2019 / Online: 28 February 2019 (11:30:13 CET)
How to cite:
Sohrab, S. S.; El-Kafrawy, S. A.; Mirza, Z.; Azhar, E. I. In silico Prediction and Cytotoxicity Evaluation of siRNAs Targeting Conserved Regions of MERS-CoV in Cell Culture. Preprints2019, 2019020268. https://doi.org/10.20944/preprints201902.0268.v1
Sohrab, S. S.; El-Kafrawy, S. A.; Mirza, Z.; Azhar, E. I. In silico Prediction and Cytotoxicity Evaluation of siRNAs Targeting Conserved Regions of MERS-CoV in Cell Culture. Preprints 2019, 2019020268. https://doi.org/10.20944/preprints201902.0268.v1
Sohrab, S. S.; El-Kafrawy, S. A.; Mirza, Z.; Azhar, E. I. In silico Prediction and Cytotoxicity Evaluation of siRNAs Targeting Conserved Regions of MERS-CoV in Cell Culture. Preprints2019, 2019020268. https://doi.org/10.20944/preprints201902.0268.v1
APA Style
Sohrab, S. S., El-Kafrawy, S. A., Mirza, Z., & Azhar, E. I. (2019). <em>In silico</em> Prediction and Cytotoxicity Evaluation of siRNAs Targeting Conserved Regions of MERS-CoV in Cell Culture. Preprints. https://doi.org/10.20944/preprints201902.0268.v1
Chicago/Turabian Style
Sohrab, S. S., Zeenat Mirza and Esam Ibraheem Azhar. 2019 "<em>In silico</em> Prediction and Cytotoxicity Evaluation of siRNAs Targeting Conserved Regions of MERS-CoV in Cell Culture" Preprints. https://doi.org/10.20944/preprints201902.0268.v1
Abstract
The Middle East Respiratory Syndrome Coronavirus (MERS-CoV) was isolated in 2012 and is well known to cause the respiratory syndrome. The orf1ab gene is known to mediate MERS-CoV replication. In this study, we have discussed the in silico prediction of potential siRNAs targeting MERS-CoV-orf1ab gene for antiviral therapeutics. To identify the potential siRNAs, various factors were considered. We have excluded the siRNAs with off-target effects and potential binding with human mRNAs. By using available softwares, total twenty-one functional, off-target reduced potential siRNA were selected from four hundred and sixty-two siRNAs based on greater potency and specificity. We have tested only seven siRNAs initially to evaluate their performance by reverse transfection approach by lipofectamine mediated delivery in Vero cells. The evaluation results showed no cytotoxicity at various concentrations of siRNAs used. The results obtained in this study provided preliminary information about the cytotoxicity which will help us to further evaluate siRNAs in other cell cultures to find out the replication inhibition efficiency of MERS-CoV. Finally, it is concluded that the in silico prediction and designing resulted in filtration and selection of potential siRNAs with high accuracy, efficiency, and strength which can be further utilized for the development of oligonucleotide-based therapeutics.
Keywords
MERS-CoV, siRNAs, in silico design, Vero cells.
Subject
Biology and Life Sciences, Virology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.