Development of murine leukemia virus-based retroviral vectors with a minimum possibility of cis-activation View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2010-10-14

AUTHORS

J Jang, J-T Lee, K Lee, S Kim, J Y Kim, K Yoon, S Kim

ABSTRACT

The possibility of insertional mutagenesis in retroviral gene therapy can be reduced by using a vector lacking the enhancer sequence present in the U3 of the long-terminal repeats. However, such vectors suffer from many pitfalls. We attempted to improve a murine leukemia virus-based retroviral vector containing the enhancer-free U3, first by making it easier to construct a producer line and then by introducing the cellular RPL10 promoter as an internal promoter. The reverse orientation of the expression cassette of the transgene was found to give higher transducing titer and higher-level gene expression. The deletion analysis revealed that the 54-bp-long sequence of U3 (34 and 20 bp present at 5′ and 3′ extreme ends, respectively) was sufficient for the functions of retroviral vectors. The data from the in vitro cell culture assay indicated that the final construct, ROK, containing all these features, had little cis-activation activity, even if it was placed right upstream from the RNA start site of the neighboring gene. Our data suggested that the newly developed vector might provide increased safety, while still producing high viral titer from a stable producer line and high-level gene expression in various target cells including human CD34+ stem cells. More... »

PAGES

240-249

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/gt.2010.135

DOI

http://dx.doi.org/10.1038/gt.2010.135

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1029482805

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/20944681


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