[High-titer preparation of HIV-1-based defective lentivector and it mediated efficient gene transfer].
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.
Zeng
Ling-Bing
LB
Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou 434000, China. zenglingbing@gmail.com
Zhang
Lin
L
Meng
Yan
Y
Yuanan
Lu
L
Ye
Lin-Bai
LB
D003673:Defective Viruses / Q000235:genetics*; D018014:Gene Transfer Techniques; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014779:Virus Replication