Baculovirus-mediated gene transfer into mammalian cells

被引:356
作者
Boyce, FM [1 ]
Bucher, NLR [1 ]
机构
[1] BOSTON UNIV,SCH MED,DEPT PATHOL,BOSTON,MA 02118
关键词
xenovector; liver; gene therapy;
D O I
10.1073/pnas.93.6.2348
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
This paper describes the use of the baculovirus Arctographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a vector for gene delivery into mammalian cells, A modified AcMNPV virus was prepared that carried the Escherichia coli lacZ reporter gene under control of the Rous sarcoma virus promoter and mammalian RNA processing signals, This modified baculovirus was then used to infect a variety of mammalian cell lines, After infection of the human liver cell lines HepG2, >25% of the cells showed high-level expression of the transduced gene, Over 70% of the cells in primary cultures of rat hepatocytes showed expression of beta-galactosidase after exposure to the virus, Cell lines from other tissues showed less or no expression of lacZ after exposure to the virus, The block to expression in less susceptible cells does not appear to result from the ability to be internalized by the target cell but rather by events subsequent to viral entry, The onset of lacZ expression occurred within 6 hr of infection in HepG2 cells and peaked 12-24 hr postinfection, Because AcMNPV is able to replicate only in insect hosts, is able to carry large (>1.5 kb) inserts, and is a highly effective gene delivery vehicle for primary cultures of hepatocytes, AcMNPV may be a useful vector for genetic manipulation of liver cells.
引用
收藏
页码:2348 / 2352
页数:5
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