Development of a packaging cell line for propagation of replication-deficient adenovirus vector.
- Author:
Jong Sik KIM
1
;
Seung Hoon LEE
;
Yong Suk CHO
;
Kyoung Sook PARK
;
Young Ho KIM
;
Je Ho LEE
Author Information
1. Clinical Research Center, Samsung Biomedical Research Institute, Seoul, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
adenovirus;
replication competent virus;
packaging cell line;
HeLa-E1 cell
- MeSH:
Adenoviridae/*genetics/physiology;
Adenovirus E1 Proteins/*genetics/metabolism;
Cell Line;
Genes, Viral;
Genes, p53;
*Genetic Vectors;
*Hela Cells;
Human;
Protein p53/genetics/metabolism;
Recombination, Genetic;
Support, Non-U.S. Gov't;
Tumor Cells, Cultured;
Virus Cultivation;
Virus Replication
- From:Experimental & Molecular Medicine
2001;33(3):145-149
- CountryRepublic of Korea
- Language:English
-
Abstract:
A human embryonic kidney cell line 293 is widely used for adenovirus production and propagation. With this cell line, however, replication-competent virus (RCV) is frequently generated, especially during large-scale production and successive propagation because 293 cells contain not only E1 gene but also non-E1 adenovirus gene. Homologous recombination between non-E1 region of 293 genomic DNA and its homologous region in the recombinant adenoviral vector generate RCV. To overcome this problem, we developed a new packaging cell line, Hela-E1, which contains minimum E1 region and from which non-E1 adenoviral region that is homologous with recombinant adenovirus vector was excluded. No RCV was detected during adenovirus propagation in Hela-E1 compared to in 293. In addition, adenovirus-p53 produced in HeLa-E1 was able to overexpress p53 protein when introduced into an ovarian cancer cell line, SKOV3. These results may have a significant impact on the development of packaging cell lines for replication-deficient adenovirus production.
