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In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs

Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention...

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Autores principales: Arani, Mohammad Javad Hajihasani, Mokhtari, Azam, Saffar, Behnaz, Asadi Samani, Leila
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8382959/
https://www.ncbi.nlm.nih.gov/pubmed/34447803
http://dx.doi.org/10.3389/fvets.2021.708591
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author Arani, Mohammad Javad Hajihasani
Mokhtari, Azam
Saffar, Behnaz
Asadi Samani, Leila
author_facet Arani, Mohammad Javad Hajihasani
Mokhtari, Azam
Saffar, Behnaz
Asadi Samani, Leila
author_sort Arani, Mohammad Javad Hajihasani
collection PubMed
description Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV- X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively. Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ~88% up to 99% in comparison with the control groups. Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV.
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spelling pubmed-83829592021-08-25 In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs Arani, Mohammad Javad Hajihasani Mokhtari, Azam Saffar, Behnaz Asadi Samani, Leila Front Vet Sci Veterinary Science Background: Border disease is believed to be one of the most important diseases in the animal husbandry industry, which has not yet been eradicated in Iran. The development of approaches based on the application of interfering RNA (RNAi) for antiviral therapy has attracted a great deal of attention over the recent years. The present research was conducted to design, construct, and apply shRNA against the NS3 gene of BDV to evaluate the prevention of BDV proliferation in the cell culture system. For this purpose, the suitable oligonucleotide sequence of NS3 gene coding was selected utilizing BDV- X818 strain. Afterwards, using shRNA design software, shRNA molecules were designed and synthesized. These shRNAs were cloned into the desired vectors and were finally transfected in HEK293T cells employing the third generation of lentiviral packaging system. Subsequently, these shRNA expressing lentiviruses were transduced to the MDBK cell line to challenge to border virus. In order to evaluate the efficacy of shRNAs, the viral infectious titer and RNA copy number were calculated with TCID50 and Real-time RT-PCR tests, respectively. Results: The results revealed that shRNAs 1, 2, and 3 decreased viral RNA by more than 90% compared to the control groups. BDV titer noticeably decreased after the challenge with shRNAs 1, 2, and 3 from ~88% up to 99% in comparison with the control groups. Conclusions: Overall, it could be concluded that RNAi may be considered as a strong treatment proposal against viruses, such as BDV. Frontiers Media S.A. 2021-08-10 /pmc/articles/PMC8382959/ /pubmed/34447803 http://dx.doi.org/10.3389/fvets.2021.708591 Text en Copyright © 2021 Arani, Mokhtari, Saffar and Asadi Samani. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Arani, Mohammad Javad Hajihasani
Mokhtari, Azam
Saffar, Behnaz
Asadi Samani, Leila
In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title_full In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title_fullStr In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title_full_unstemmed In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title_short In vitro Inhibition of Border Disease Virus Replication With Lentivirus-Mediated shRNAs
title_sort in vitro inhibition of border disease virus replication with lentivirus-mediated shrnas
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8382959/
https://www.ncbi.nlm.nih.gov/pubmed/34447803
http://dx.doi.org/10.3389/fvets.2021.708591
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