Development of Versatile and Flexible Sf9 Packaging Cell Line-Dependent OneBac System for Large-Scale Recombinant Adeno-Associated Virus Production

doi:10.1089/hgtb.2019.123

	Development of Versatile and Flexible Sf9 Packaging Cell Line-Dependent OneBac System for Large-Scale Recombinant Adeno-Associated Virus Production
	Wu, Yang; Mei, Ting; Jiang, Liangyu; Han, Zengpeng; Dong, Ruping; Yang, Tian; Xu, Fudiang
刊名	HUMAN GENE THERAPY METHODS
	2019-10-01
卷号	30 期号:5 页码:172-183
关键词	rAAV production OneBac recombinant baculovirus Sf9 packaging cell line gene therapy
ISSN号	1946-6536
DOI	10.1089/hgtb.2019.123
英文摘要	Recombinant adeno-associated viruses (rAAVs) are excellent vectors for gene delivery. However, current Sf9/Cap-Rep packaging cell line-dependent OneBac systems still lack versatility and flexibility for large-scale production of rAAVs. In this study, we developed an improved OneBac system that includes a novel dual-function baculovirus expression vector (BEV) termed BEV/Cap-(ITR-GOI) that carries both the AAV Cap gene and rAAV genome inverted terminal repeat (ITR) sequences flanking the gene of interest (GOI), a versatile Sf9-GFP/Rep packaging cell line that harbors silent copies of the AAV2 Rep gene that can be expressed after BEV infection, and constitutively expressed green fluorescent protein (GFP) reporter genes to facilitate cell line screening. The BEV/Cap-(ITR-GOI) construct allows flexibility to switch among different Cap gene serotypes using simple BEV reconstruction, and is stable for at least five serial passages. Furthermore, the Sf9-GFP/Rep stable cell line is versatile for production of different rAAV serotypes. The yield levels for rAAV2, rAAV8, and rAAV9 exceeded 10(5) vector genomes (VG) per cell, which is similar to other currently available large-scale rAAV production systems. The new Bac system-derived rAAVs have biophysical properties similar to HEK293 cell-derived rAAVs, as well as high quality and activity. In summary, the novel Sf9-GFP/Rep packaging cell line-dependent OneBac system can facilitate large-scale rAAV production and rAAV-based gene therapy.
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] ; State Key Program of National Natural Science Foundation of China[31830035] ; Strategic Priority Research Program (B)[XDB0205005]
WOS关键词	INSECT CELLS ; MINIMAL ENCAPSIDATION ; VIRAL VECTORS ; GENE-TRANSFER ; RAAV VECTORS ; PURIFICATION ; EXPRESSION ; TITER ; REP
WOS研究方向	Biotechnology & Applied Microbiology ; Genetics & Heredity ; Research & Experimental Medicine
语种	英语
出版者	MARY ANN LIEBERT, INC
WOS记录号	WOS:000490390300002
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 ; State Key Program of National Natural Science Foundation of China ; State Key Program of National Natural Science Foundation 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 ; State Key Program of National Natural Science Foundation of China ; State Key Program of National Natural Science Foundation 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 ; State Key Program of National Natural Science Foundation of China ; State Key Program of National Natural Science Foundation 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 ; State Key Program of National Natural Science Foundation of China ; State Key Program of National Natural Science Foundation 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/112942/21403]
专题	中国科学院武汉物理与数学研究所
通讯作者	Wu, Yang; Xu, Fudiang
作者单位	Chinese Acad Sci, Ctr Excellence Brain Sci & Intelligent Technol, Wuhan Inst Phys & Math,State Key Lab Magnet Reson, Key Lab Magnet Resonance Biol Syst,Wuhan Ctr Magn, Wuhan, Hubei, Peoples R China
推荐引用方式 GB/T 7714	Wu, Yang,Mei, Ting,Jiang, Liangyu,et al. Development of Versatile and Flexible Sf9 Packaging Cell Line-Dependent OneBac System for Large-Scale Recombinant Adeno-Associated Virus Production[J]. HUMAN GENE THERAPY METHODS,2019,30(5):172-183.
APA	Wu, Yang.,Mei, Ting.,Jiang, Liangyu.,Han, Zengpeng.,Dong, Ruping.,...&Xu, Fudiang.(2019).Development of Versatile and Flexible Sf9 Packaging Cell Line-Dependent OneBac System for Large-Scale Recombinant Adeno-Associated Virus Production.HUMAN GENE THERAPY METHODS,30(5),172-183.
MLA	Wu, Yang,et al."Development of Versatile and Flexible Sf9 Packaging Cell Line-Dependent OneBac System for Large-Scale Recombinant Adeno-Associated Virus Production".HUMAN GENE THERAPY METHODS 30.5(2019):172-183.