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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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/9/1/164 |
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| author | Arjun Challagulla Kristie A. Jenkins Terri E. O’Neil Shunning Shi Kirsten R. Morris Terry G. Wise Prasad N. Paradkar Mark L. Tizard Timothy J. Doran Karel A. Schat |
| author_facet | Arjun Challagulla Kristie A. Jenkins Terri E. O’Neil Shunning Shi Kirsten R. Morris Terry G. Wise Prasad N. Paradkar Mark L. Tizard Timothy J. Doran Karel A. Schat |
| author_sort | Arjun Challagulla |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T04:56:33Z |
| format | Article |
| id | doaj.art-c9ca6b57a53f40fe882f9d484bfc93f7 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T04:56:33Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-c9ca6b57a53f40fe882f9d484bfc93f72023-12-03T13:05:08ZengMDPI AGMicroorganisms2076-26072021-01-019116410.3390/microorganisms9010164In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4Arjun Challagulla0Kristie A. Jenkins1Terri E. O’Neil2Shunning Shi3Kirsten R. Morris4Terry G. Wise5Prasad N. Paradkar6Mark L. Tizard7Timothy J. Doran8Karel A. Schat9CSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaCSIRO Health and Biosecurity, Australian Centre for Disease Preparedness, Geelong, VIC 3220, AustraliaDepartment of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USAMarek’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.https://www.mdpi.com/2076-2607/9/1/164Marek’s disease virusCRISPR/Cas9ICP4transgenic chickendisease resilience |
| spellingShingle | Arjun Challagulla Kristie A. Jenkins Terri E. O’Neil Shunning Shi Kirsten R. Morris Terry G. Wise Prasad N. Paradkar Mark L. Tizard Timothy J. Doran Karel A. Schat In Vivo Inhibition of Marek’s Disease Virus in Transgenic Chickens Expressing Cas9 and gRNA against ICP4 Microorganisms Marek’s disease virus CRISPR/Cas9 ICP4 transgenic chicken disease resilience |
| 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 | Marek’s disease virus CRISPR/Cas9 ICP4 transgenic chicken disease resilience |
| url | https://www.mdpi.com/2076-2607/9/1/164 |
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