A Recombinant Baculovirus Efficiently Generates Recombinant Adeno-Associated Virus Vectors in Cultured Insect Cells and Larvae

doi:10.1016/j.omtm.2018.05.005

	A Recombinant Baculovirus Efficiently Generates Recombinant Adeno-Associated Virus Vectors in Cultured Insect Cells and Larvae
	Xu, Fuqiang 1,2,3; Lin, Kunzhang 2,3; He, Xiaobing 2,3; Han, Zengpeng 2,3; Yang, Tian 2,3; Geng, Hao 2,3; Jiang, Liangyu 2,3; Wu, Yang 2,3
刊名	MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
	2018-09-01
卷号	10 页码:38-47
ISSN号	2329-0501
DOI	10.1016/j.omtm.2018.05.005
英文摘要	Current large-scale recombinant adeno-associated virus (rAAV) production systems based on the baculovirus expression vector (BEV) remain complicated and cost-intensive, and they lack versatility and flexibility. Here we present a novel recombinant baculovirus integrated with all packaging elements for the production of rAAV. To optimize BEV construction, ribosome leaky-scanning mechanism was used to express AAV Rep and Cap proteins downstream of the PH and P10 promoters in the pFast.Bac.Dual vector, respectively, and the rAAV genome was inserted between the two promoters. The yields of rAAV2, rAAV8, and rAAV9 derived from the BEV-infected Sf9 cells exceeded 10(5) vector genomes (VG) per cell. The BEV was shown to be stable and showed no apparent decrease of rAAV yield after at least four serial passages. The rAAVs derived from the new Bac system displayed high-quality and high-transduction activity. Additionally, rAAV2 could be efficiently generated from BEV-infected beet armyworm larvae at a per-larvae yield of 2.75 +/- 1.66 x 10(10) VG. The rAAV2 derived from larvae showed a structure similar to the rAAV2 derived from HEK293 cells, and it also displayed high-transduction activity. In summary, the novel BEV is ideally suitable for large-scale rAAV production. Further, this study exploits a potential cost-efficient platform for rAAV production in insect larvae.
资助项目	National Natural Science Foundation of China[31500868] ; National Natural Science Foundation of China[31771156] ; National Basic Research Program (973 Program) of China[2015CB755600] ; Strategic Priority Research Program (B)[XDB0205005]
WOS关键词	GENE-TRANSFER ; MINIMAL ENCAPSIDATION ; VIRAL VECTORS ; RAAV VECTORS ; AAV VECTORS ; EXPRESSION ; PURIFICATION ; SYSTEM ; NUCLEOPOLYHEDROVIRUS ; TITER
WOS研究方向	Research & Experimental Medicine
语种	英语
出版者	CELL PRESS
WOS记录号	WOS:000451115100005
资助机构	National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Basic Research Program (973 Program) of China ; National Basic Research Program (973 Program) of China ; Strategic Priority Research Program (B) ; Strategic Priority Research Program (B) ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Basic Research Program (973 Program) of China ; National Basic Research Program (973 Program) of China ; Strategic Priority Research Program (B) ; Strategic Priority Research Program (B) ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Basic Research Program (973 Program) of China ; National Basic Research Program (973 Program) of China ; Strategic Priority Research Program (B) ; Strategic Priority Research Program (B) ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Basic Research Program (973 Program) of China ; National Basic Research Program (973 Program) of China ; Strategic Priority Research Program (B) ; Strategic Priority Research Program (B)
内容类型	期刊论文
源URL	[http://ir.wipm.ac.cn/handle/112942/13522]
专题	中国科学院武汉物理与数学研究所
通讯作者	Xu, Fuqiang; Wu, Yang
作者单位	1.Chinese Acad Sci, Ctr Excellence Brain Sci & Intelligence Technol, Shanghai 200031, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Phys & Math, Brain Res Ctr, Wuhan 430071, Hubei, Peoples R China 3.Chinese Acad Sci, Wuhan Inst Phys & Math, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan 430071, Hubei, Peoples R China
推荐引用方式 GB/T 7714	Xu, Fuqiang,Lin, Kunzhang,He, Xiaobing,et al. A Recombinant Baculovirus Efficiently Generates Recombinant Adeno-Associated Virus Vectors in Cultured Insect Cells and Larvae[J]. MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT,2018,10:38-47.
APA	Xu, Fuqiang.,Lin, Kunzhang.,He, Xiaobing.,Han, Zengpeng.,Yang, Tian.,...&Wu, Yang.(2018).A Recombinant Baculovirus Efficiently Generates Recombinant Adeno-Associated Virus Vectors in Cultured Insect Cells and Larvae.MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT,10,38-47.
MLA	Xu, Fuqiang,et al."A Recombinant Baculovirus Efficiently Generates Recombinant Adeno-Associated Virus Vectors in Cultured Insect Cells and Larvae".MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT 10(2018):38-47.