Efficient protein expression from the endogenous RNA polymerase I promoter using a human ribosomal DNA targeting vector

Vector systems to deliver, integrate and express therapeutic genes in host cells are essential for gene therapy. In the present study, we investigated a novel vector system for integration and expression of a transgene. In this system, the transgene expression was driven by an endogenous RNA polymer...

Full description

Saved in:
Bibliographic Details
Published in:Biochemical and biophysical research communications 2008-03, Vol.367 (4), p.846-851
Main Authors: Wen, Lu, Liang, De Sheng, Wu, Ling Qian, Cai, Fang, Pan, Qian, Long, Zhi Gao, Dai, He Ping, Xia, Kun, Xia, Jia Hui
Format: Article
Language:English
Subjects:
Cell Line, Tumor
Coagulation factor IX
DNA, Ribosomal - genetics
Factor IX - biosynthesis
Factor IX - genetics
Fibrosarcoma - genetics
Fibrosarcoma - metabolism
Gene expression
Gene targeting
Gene Targeting - methods
Gene therapy
Genetic Vectors - genetics
Homologous recombination
Humans
Non-viral
Promoter Regions, Genetic - genetics
Protein Engineering - methods
Recombinant Proteins - metabolism
Ribosomal DNA
RNA polymerase I
RNA Polymerase I - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Vector systems to deliver, integrate and express therapeutic genes in host cells are essential for gene therapy. In the present study, we investigated a novel vector system for integration and expression of a transgene. In this system, the transgene expression was driven by an endogenous RNA polymerase I (Pol I) promoter after being integrated into the ribosomal DNA (rDNA) locus. Human coagulation factor IX coding sequence (FIX), with an internal ribosome entry sites element at its leader region, was targeted into the 18S rDNA locus via homologous recombination. FIX protein expression, which was under the control of the endogenous Pol I promoter, was found to be similar to that of a moderate Pol II promoter. The average FIX expression level of the rDNA recombinants was additionally enhanced to that from a strong Pol II promoter as a result of elimination of position effects. Our data suggest the possibility of applying this system in gene therapy for hereditary diseases.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2008.01.018