Construction of circular RNA vaccine against porcine reproductive and respiratory syndrome virus and optimization of circularization conditions
10.16303/j.cnki.1005-4545.2025.05.02
- VernacularTitle:猪繁殖与呼吸综合征病毒环状RNA疫苗的构建及环化条件优化
- Author:
Chunhao TAO
1
;
Ying HUANG
;
Zhen WANG
;
Yitong JIANG
;
Hongfei ZHU
;
Hong JIA
Author Information
1. 中国农业科学院北京畜牧兽医研究所,北京 100193
- Publication Type:Journal Article
- Keywords:
circRNA vaccine;
preparation system;
PRRSV vaccine;
in vitro expression
- From:
Chinese Journal of Veterinary Science
2025;45(5):896-904
- CountryChina
- Language:Chinese
-
Abstract:
In order to establish a highly efficient,convenient,and effective circular RNA(circRNA)vaccine preparation system,enhanced green fluorescent protein(EGFP)circRNA was constructed using permuted intron exon(PIE)strategy based on type Ⅰ introns.Then,circRNA circularization rates of RNA after in vitro transcription(IVT),primary circularization(IVC1),and secondary circularization(IVC2)were compared after purification.The constructed circRNA system was fur-ther applied to porcine reproductive and respiratory syndrome virus(PRRSV),and two circRNAs based on PRRSV GP5 protein were constructed and developed for in vitro expression.Results showed that circularization rates and protein expression effects of EGFP circRNA in IVC1 RNA and IVC2 RNA were similar,but both were significantly better than those of IVT RNA.Purity of EGFP circRNA reached 74％,and purities of two PRRSV GP5 protein circRNAs constructed using this preparation system were 71％and 64％,respectively.Western blot and indirect immunofluo-rescence assay(IFA)results indicated that both of the PRRSV GP5 protein circRNAs were suc-cessfully expressed.The results demonstrated that an easy-to-operate,low-cost circRNA prepara-tion system with high circularization rate was successfully constructed.Two PRRSV GP5 protein circRNA vaccines were successfully prepared using this system and expressed efficiently,which provides a reference for the development of animal circRNA vaccines and novel candidate vaccines against PRRSV.
