Introduction of DNA into rat liver with a hand-held gene gun: distribution of the expressed enzyme, [32P]DNA, and Ca2+ flux.
DNA-coated Au particles were accelerated by pressurized He gas to supersonic velocities for introduction of a gene into cells. Experimental and theoretical analyses both revealed a heterogeneous distribution of the particles per shot (1 mg Au = 2.4 x 10(7) particles with 2 microg [32P] DNA = 2.5 x 10(11) moles). For introduction of genes into the liver of living rats, the best results were obtained with a newly developed hand-held gene delivery system. The beta-galactosidase gene introduced into rat liver with Au particles by He at 250 psi was expressed (1.2 microunits/microg protein) in a limited area of the liver surface (8 x 8 mm, depth 0.5 mm). When the same gene gun was used on a monolayer of cultured COS7 cells (about 5 microm thick), cells were lost in the central area of heavy bombardment. Cell death caused by influx of Ca2+ was prevented by the use of the cytosol-type culture medium.
Yoshida
Y
Y
Department of Gastroenterology, Jichi Omiya Medical Center, Japan.
Kobayashi
E
E
Endo
H
H
Hamamoto
T
T
Yamanaka
T
T
Fujimura
A
A
Kagawa
Y
Y
D000818:Animals; D019470:Biolistics; D019556:COS Cells; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage*; D006046:Gold / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D051381:Rats; D001616:beta-Galactosidase / Q000378:metabolism
D010761:Phosphorus Radioisotopes; D006046:Gold; D004247:DNA; D001616:beta-Galactosidase; D002118:Calcium