Gene Transfer into Mammalian Cells Using Histone-Condensed Plasmid DNA

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Abstract
A recombinant histone (NLS-H1) containing both the SV40 large T antigen nuclear localization signal and the carboxy-terminal domain of human histone H1° was produced in bacteria. NLS-H1–plasmid DNA complexes, in the presence of chloroquine, mediated reporter gene transfer into cultured cells with similar efficiencies as plasmid DNA–cationic lipid (lipofectin) complexes. NIH-3T3 or COS-7 cells transfected with NLS-H1–plasmid DNA–lipofectin complexes expressed at least 20 times more luciferase or had at least 2.5 times more β-galactosidase-positive cells than those transfected with plasmid DNA–lipofectin complexes. Foreign gene expression was also improved by other DNA-binding proteins and cationic lipid formulations, yet the greatest enhancement was obtained with complexes containing either NLS-H1 or calf thymus histone H1. Histone H1-plasmid DNA–lipofectin complexes were internalized by a greater number of cells than plasmid DNA–lipofectin complexes. Both cationic lipid- and receptor-mediated gene transfer techniques have been successful in delivering genes to eukaryotic cells in culture but have been limited in their application in vivo. A recombinant histone was developed to study the influence of DNA condensation upon gene delivery. Results indicated that recombinant histone–DNA complexes successfully transferred genes in vitro in the presence of endosomolytic compounds. Enhancement in gene delivery and expression was observed following the addition of recombinant histone to cationic lipid–DNA complexes.