Plat-E: an efficient and stable system for transient packaging of retroviruses View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2000-06

AUTHORS

S Morita, T Kojima, T Kitamura

ABSTRACT

A potent retrovirus packaging cell line named Platinum-E (Plat-E) was generated based on the 293T cell line. Plat-E is superior to existing packaging cell lines regarding efficiency, stability and safety. The novel packaging constructs utilized in establishment of Plat-E ensure high and stable expression of viral structural proteins. Conventional packaging constructs made use of the promoter of MuLV-LTR for expression of viral structural genes gag-pol and env, while our packaging constructs utilized the EF1alpha promoter, which is 100-fold more potent than the MuLV-LTR in 293T cells in combination with the Kozak's consensus sequence upstream of the initiation codon resulting in high expression of virus structural proteins in Plat-E cells. To maintain the high titers of retroviruses under drug selection pressure, we inserted the IRES (internal ribosome entry site) sequence between the gene encoding gag-pol or env, and the gene encoding a selectable marker in the packaging constructs. Plat-E cells can stably produce retroviruses with an average titer of 1 x 107/ml for at least 4 months. In addition, as we used only the coding sequences of viral structural genes to avoid inclusion of unnecessary retrovirus sequences in the packaging constructs, the probability of generating the replication competent retroviruses (RCR) by recombination can virtually be ruled out. More... »

PAGES

3301206

Journal

TITLE

Gene Therapy

ISSUE

12

VOLUME

7

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/sj.gt.3301206

DOI

http://dx.doi.org/10.1038/sj.gt.3301206

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1032864882

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/10871756


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