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.
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.
复制标题
DOI：
10.3390/vaccines10050686
复制
发表时间：
2022-04-27
期刊：
影响因子：
7.8
通讯作者：
Sthitmatee, Nattawooti
中科院分区：
文献类型：
--
作者：
Apinda, Nisachon;Yao, Yongxiu;Zhang, Yaoyao;Reddy, Vishwanatha R. A. P.;Chang, Pengxiang;Nair, Venugopal;Sthitmatee, Nattawooti
鸭肠炎病毒（DEV）和多杀性巴氏杆菌是鸭瘟和禽霍乱的病原体，是鸭的急性传染病，是导致鸭发病和死亡的主要原因。本研究采用NHEJ-CRISPR/Cas9介导的基因编辑策略，配合Cre-Lox系统，表明DEV Jansen减毒株基因组中UL55-LORF11和UL44-44.5位点的两个新位点可用于多杀性巴氏杆菌外膜蛋白H（ompH）基因的稳定表达，可作为具有保护鸭子潜力的二价候选疫苗对抗主要病毒和细菌病原体。与野生型病毒相比，分别在 UL55-LORF11 和 UL44-44.5 位点插入 ompH-V5 基因的两种重组病毒 DEV-OmpH-V5-UL55-LORF11 和 DEV-OmpH-V5-UL44-44.5 表现出相似的生长动力学和噬菌斑大小，证实插入外源基因不会对病毒产生任何有害影响。开发。这是首次应用CRISPR/Cas9系统将来自细菌的高免疫原性基因快速有效地插入DEV基因组中。这种方法提供了一种将其他抗原引入 DEV 基因组的多价载体的有效方法。
Duck enteritis virus (DEV) and Pasteurella multocida, 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 (ompH) gene of P. multocida 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.
登录
查看更多内容
影响因子：
4.6
作者：
Liang X;Sun L;Yu T;Pan Y;Wang D;Hu X;Fu Z;He Q;Cao G
通讯作者：
Cao G
将腺病毒载体用作兽医疫苗。
影响因子：
5.1
作者：
Ferreira TB;Alves PM;Aunins JG;Carrondo MJ
通讯作者：
Carrondo MJ
影响因子：
1.2
作者：
Sthitmatee, Nattawooti;Yano, Terdsak;Sawada, Takuo
通讯作者：
Sawada, Takuo
DOI：
10.3390/v10020081
发表时间：
2018-02-13
期刊：
Viruses
影响因子：
--
作者：
Chang P;Yao Y;Tang N;Sadeyen JR;Sealy J;Clements A;Bhat S;Munir M;Bryant JE;Iqbal M
通讯作者：
Iqbal M
DOI：
10.1292/jvms.17-0176
发表时间：
2018-01-27
期刊：
The Journal of veterinary medical science
影响因子：
--
作者：
Muangthai K;Tankaew P;Varinrak T;Uthi R;Rojanasthien S;Sawada T;Sthitmatee N
通讯作者：
Sthitmatee N