title: [High-titer preparation of HIV-1-based defective lentivector and it mediated efficient gene transfer].
abstract: Lentivectors have drawn considerable attention recently and become important delivery vehicles for gene transfer manipulation. By Transiently co-transfecting 293T packaging cells with three DNA plasmids system encoding lentivector constituents, a protocol for bulky preparation of human immunodeficiency virus type-1 ( HIV-1)-based defective lentivector with high titer has been established. Transient co-transfection of 293T packaging cells resulted in production of high-titer vector (1.1 x 10 IU/ml), which can be further concentrated over 100-fold through a single step centrifugation. The vector was capable of efficiently transducing a variety of cells from both primate and non-primate sources, including of human T-lymphoblastoid cell line. Long-term culture of vector transduced cells showed a stable expression of foreign gene over 18 months detected by RT-PCR. Assessment of potential generation of replication-competent virus revealed no detection of p24 antigen protein or infectious particles in vector-transduced cells.
authors.affiliation: Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou 434000, China. zenglingbing@gmail.com
authors.forename: Ling-Bing
authors.initials: LB
authors.lastname: Zeng
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Meng
authors.forename: Lu
authors.initials: L
authors.lastname: Yuanan
authors.forename: Lin-Bai
authors.initials: LB
authors.lastname: Ye
mesh_terms: D003673:Defective Viruses / Q000235:genetics*; D018014:Gene Transfer Techniques; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014779:Virus Replication