CRISPR/Cas9 Editing of Duck Enteritis Virus Genome for the Construction of a Recombinant Vaccine Vector Expressing ompH Gene of Pasteurella multocida in Two Novel Insertion Sites

Duck enteritis virus (DEV) and , the causative agent of duck plague and fowl cholera, are acute contagious diseases and leading causes of morbidity and mortality in duck. The NHEJ-CRISPR/Cas9-mediated gene editing strategy, accompanied with the Cre-Lox system, have been employed in the present study...

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Bibliographic Details
Published in:Vaccines (Basel) 2022-04, Vol.10 (5), p.686
Main Authors: Apinda, Nisachon, Yao, Yongxiu, Zhang, Yaoyao, Reddy, Vishwanatha R A P, Chang, Pengxiang, Nair, Venugopal, Sthitmatee, Nattawooti
Format: Article
Language:English
Subjects:
Antigens
Aquatic birds
Artificial chromosomes
Cell cycle
Cholera
Cloning
Cre-Lox
CRISPR
CRISPR-Cas9
duck enteritis virus
Duck plague
Embryos
Enteritis
Fowl cholera
Genetic modification
Genomes
Growth kinetics
H protein
Immunogenicity
Infections
Insertion
Liquid oxygen
Loci
Membrane proteins
Morbidity
NHEJ
Non-homologous end joining
Pasteurella multocida
Plague
Proteins
Vaccines
Veterinary medicine
Viruses
Waterborne diseases
Waterfowl
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Summary:Duck enteritis virus (DEV) and , the causative agent of duck plague and fowl cholera, are acute contagious diseases and leading causes of morbidity and mortality in duck. The NHEJ-CRISPR/Cas9-mediated gene editing strategy, accompanied with the Cre-Lox system, have been employed in the present study to show that two new sites at UL55-LORF11 and UL44-44.5 loci in the genome of the attenuated Jansen strain of DEV can be used for the stable expression of the outer membrane protein H ( ) gene of that could be used as a bivalent vaccine candidate with the potential of protecting ducks simultaneously against major viral and bacterial pathogens. The two recombinant viruses, DEV-OmpH-V5-UL55-LORF11 and DEV-OmpH-V5-UL44-44.5, with the insertion of ompH-V5 gene at the UL55-LORF11 and UL44-44.5 loci respectively, showed similar growth kinetics and plaque size, compared to the wildtype virus, confirming that the insertion of the foreign gene into these did not have any detrimental effects on DEV. This is the first time the CRISPR/Cas9 system has been applied to insert a highly immunogenic gene from bacteria into the DEV genome rapidly and efficiently. This approach offers an efficient way to introduce other antigens into the DEV genome for multivalent vector.
ISSN:2076-393X
2076-393X
DOI:10.3390/vaccines10050686