Gene transfer into mammalian cells using histone-condensed plasmid DNA.
A recombinant histone (NLS-H1) containing both the SV40 large T antigen nuclear localization signal and the carboxy-terminal domain of human histone H1(0) 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 beta-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.
Fritz
J D
JD
Department of Pediatrics and Medical Genetics, Waisman Center, University of Wisconsin, Madison 53705, USA.
Herweijer
H
H
Zhang
G
G
Wolff
J A
JA
D016475:3T3 Cells; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000502:physiology; D019556:COS Cells; D002417:Cattle; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002738:Chloroquine / Q000494:pharmacology; D004274:DNA, Recombinant / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000648:ultrastructure; D006367:HeLa Cells; D006657:Histones / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins
D000952:Antigens, Polyomavirus Transforming; D004274:DNA, Recombinant; D006657:Histones; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins; C020888:1,2-dielaidoylphosphatidylethanolamine; D002738:Chloroquine; D008156:Luciferases