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Construction of an EGF receptor-mediated histone H1(0)-based gene delivery system
Fei-Han Dai1, Yan Chen, Chang-Chun Ren
1National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, 25/Ln 2200, Xie Tu Road, 200032, Shanghai, P.R. China.
Purpose:
To construct an EGF receptor (EGF-R)-mediated histone H1(0)-based gene delivery system for gene therapy.
Methods:
A recombinant DNA containing histone H1(0), EGF-R ligand, and endosomalytic domains was constructed in a prokaryotic vector and expressed in E. coli. Expression of the beta-galactosidase (beta-gal) gene in the tumor cells and tissues was observed after transduction of the beta-gal gene packaged by purified fusion proteins in vitro and in vivo.
Results:
As an extension of the research on previously reported chemically synthetic composite polypeptide gene delivery systems, this genetically engineered polypeptide has proved to be capable of targeting the beta-galactosidase (beta-gal) gene into EGF-R-positive cancer cells both in vitro and in vivo. We also studied the time course of beta-gal gene expression in tumor tissues delivered in vivo by this polypeptide vector. At 24 h after administration, expression of the beta-galactosidase gene in tumor reached peak levels. The dosage optimization of administered polyplex was also investigated. The optimal dose of polyplex per mouse was 1 microg DNA packaged by 3 microg of composite polypeptide.
Conclusions:
The genetically engineered polypeptide based on histone H1(0) is a promising gene delivery system targeting EGF-R.