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In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4

Marek’s disease (MD), caused by MD herpesvirus (MDV), is an economically important disease in chickens. The efficacy of the existing vaccines against evolving virulent stains may become limited and necessitates the development of novel antiviral strategies to protect poultry from MDV strains with in...

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Autores principales: Challagulla, Arjun, Jenkins, Kristie A., O’Neil, Terri E., Shi, Shunning, Morris, Kirsten R., Wise, Terry G., Paradkar, Prasad N., Tizard, Mark L., Doran, Timothy J., Schat, Karel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828426/
https://www.ncbi.nlm.nih.gov/pubmed/33450980
http://dx.doi.org/10.3390/microorganisms9010164
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author Challagulla, Arjun
Jenkins, Kristie A.
O’Neil, Terri E.
Shi, Shunning
Morris, Kirsten R.
Wise, Terry G.
Paradkar, Prasad N.
Tizard, Mark L.
Doran, Timothy J.
Schat, Karel A.
author_facet Challagulla, Arjun
Jenkins, Kristie A.
O’Neil, Terri E.
Shi, Shunning
Morris, Kirsten R.
Wise, Terry G.
Paradkar, Prasad N.
Tizard, Mark L.
Doran, Timothy J.
Schat, Karel A.
author_sort Challagulla, Arjun
collection PubMed
description Marek’s disease (MD), caused by MD herpesvirus (MDV), is an economically important disease in chickens. The efficacy of the existing vaccines against evolving virulent stains may become limited and necessitates the development of novel antiviral strategies to protect poultry from MDV strains with increased virulence. The CRISPR/Cas9 system has emerged as a powerful genome editing tool providing an opportunity to develop antiviral strategies for the control of MDV infection. Here, we characterized Tol2 transposon constructs encoding Cas9 and guide RNAs (gRNAs) specific to the immediate early infected-cell polypeptide-4 (ICP4) of MDV. We generated transgenic chickens that constitutively express Cas9 and ICP4-gRNAs (gICP4) and challenged them via intraabdominal injection of MDV-1 Woodlands strain passage-19 (p19). Transgenic chickens expressing both gRNA/Cas9 had a significantly reduced replication of MDV in comparison to either transgenic Cas9-only or the wild-type (WT) chickens. We further confirmed that the designed gRNAs exhibited sequence-specific virus interference in transgenic chicken embryo fibroblast (CEF) expressing Cas9/gICP4 when infected with MDV but not with herpesvirus of turkeys (HVT). These results suggest that CRISPR/Cas9 can be used as an antiviral approach to control MDV infection in chickens, allowing HVT to be used as a vector for recombinant vaccines.
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spelling pubmed-78284262021-01-25 In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4 Challagulla, Arjun Jenkins, Kristie A. O’Neil, Terri E. Shi, Shunning Morris, Kirsten R. Wise, Terry G. Paradkar, Prasad N. Tizard, Mark L. Doran, Timothy J. Schat, Karel A. Microorganisms Article Marek’s disease (MD), caused by MD herpesvirus (MDV), is an economically important disease in chickens. The efficacy of the existing vaccines against evolving virulent stains may become limited and necessitates the development of novel antiviral strategies to protect poultry from MDV strains with increased virulence. The CRISPR/Cas9 system has emerged as a powerful genome editing tool providing an opportunity to develop antiviral strategies for the control of MDV infection. Here, we characterized Tol2 transposon constructs encoding Cas9 and guide RNAs (gRNAs) specific to the immediate early infected-cell polypeptide-4 (ICP4) of MDV. We generated transgenic chickens that constitutively express Cas9 and ICP4-gRNAs (gICP4) and challenged them via intraabdominal injection of MDV-1 Woodlands strain passage-19 (p19). Transgenic chickens expressing both gRNA/Cas9 had a significantly reduced replication of MDV in comparison to either transgenic Cas9-only or the wild-type (WT) chickens. We further confirmed that the designed gRNAs exhibited sequence-specific virus interference in transgenic chicken embryo fibroblast (CEF) expressing Cas9/gICP4 when infected with MDV but not with herpesvirus of turkeys (HVT). These results suggest that CRISPR/Cas9 can be used as an antiviral approach to control MDV infection in chickens, allowing HVT to be used as a vector for recombinant vaccines. MDPI 2021-01-13 /pmc/articles/PMC7828426/ /pubmed/33450980 http://dx.doi.org/10.3390/microorganisms9010164 Text en © 2021 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Challagulla, Arjun
Jenkins, Kristie A.
O’Neil, Terri E.
Shi, Shunning
Morris, Kirsten R.
Wise, Terry G.
Paradkar, Prasad N.
Tizard, Mark L.
Doran, Timothy J.
Schat, Karel A.
In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title_full In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title_fullStr In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title_full_unstemmed In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title_short In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4
title_sort in vivo inhibition of marek’s disease virus in transgenic chickens expressing cas9 and grna against icp4
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828426/
https://www.ncbi.nlm.nih.gov/pubmed/33450980
http://dx.doi.org/10.3390/microorganisms9010164
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