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Generering af rekombinant aviær Herpesvirus vektorer med CRISPR/Cas9 gen redigering
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면역학 및 감염병학
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JoVE 신문 면역학 및 감염병학
Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

Generering af rekombinant aviær Herpesvirus vektorer med CRISPR/Cas9 gen redigering

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

자동 번역

자동 번역

Herpesvirus af kalkuner (HVT) er udbredt som en vektor platform for generation af rekombinante vacciner mod en række fjerkræsygdomme. Denne artikel beskriver en enkel og hurtig tilgang for generation af rekombinant HVT-vektoriserede vacciner ved hjælp af en integreret NHEJ-CRISPR/Cas9, og Cre-Lox system.

Tags

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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