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Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

DOI:
10.3791/58193-v

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12:21 min

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January 07, 2019

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, , , , , , , ,
1The Pirbright Institute & UK-China Centre of Excellence for Research on Avian Diseases, 2Binzhou Animal Science and Veterinary Medicine Academy & UK-China Centre of Excellence for Research on Avian Diseases

Chapters

  • 00:04Title
  • 00:53CRISPR/Cas9-mediated Knock-in: Transfection and Infection
  • 02:14Harvesting and Purification of the HVT Recombinant Virus
  • 05:38Excision of the Fluorescent Reporter Gene via the Cre-lox System
  • 06:23Plaque Purification
  • 07:31Verification of the Recombinant HVT
  • 09:31Stability of the Recombinant Virus
  • 10:36Results: Analysis of the Generated Recombinant Avian Herpesvirus Vectors
  • 11:38Conclusion

Summary

Automatic Translation

Automatic Translation

Herpesvirus of turkeys (HVT) is widely used as a vector platform for the generation of recombinant vaccines against a number of avian diseases. This article describes a simple and rapid approach for the generation of recombinant HVT-vectored vaccines using an integrated NHEJ-CRISPR/Cas9 and Cre-Lox system.

Tags

Keywords: CRISPR/Cas9Recombinant Avian HerpesvirusViral AntigensHVT GenomeRecombinant VaccinesGFP Expression CassetteCre-LoxP SystemCell SortingChick Embryo FibroblastsTransfectionHVT Virus StockPlaque Forming UnitsTrypsin-EDTACell SuspensionCell DensityCell SorterGFP-expressing Cells

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