Abstract
This paper describes the use of the baculovirus Autographa 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 (>15 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.
Dates
Type | When |
---|---|
Created | 23 years, 1 month ago (July 26, 2002, 10:36 a.m.) |
Deposited | 3 years, 4 months ago (April 13, 2022, 2:53 p.m.) |
Indexed | 3 weeks, 4 days ago (Aug. 7, 2025, 4:57 p.m.) |
Issued | 29 years, 5 months ago (March 19, 1996) |
Published | 29 years, 5 months ago (March 19, 1996) |
Published Online | 29 years, 5 months ago (March 19, 1996) |
Published Print | 29 years, 5 months ago (March 19, 1996) |
@article{Boyce_1996, title={Baculovirus-mediated gene transfer into mammalian cells.}, volume={93}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.93.6.2348}, DOI={10.1073/pnas.93.6.2348}, number={6}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Boyce, F M and Bucher, N L}, year={1996}, month=mar, pages={2348–2352} }