Effect of solution composition of plasmid DNA on gene transfection following liver surface administration in mice.
We investigated the effect of plasmid DNA (pDNA) solution composition on gene transfection following liver surface administration in mice. Gene transfection experiments in situ and in vivo were performed using the following pDNA solutions: dextrose solution, NaCl solution, phosphate buffer, phosphate-buffered saline, Tris/HCl buffer with EDTA, Tris/HCl buffer with EDTA and Triton X-100, and water. In in situ experiments, we used a glass cylindrical diffusion cell that limited the contact area between the liver surface and the naked pDNA solution. The gene transfection at the site of diffusion cell attachment increased in hypotonic solution, and decreased in hypertonic solution, compared with isotonic solution. In in vivo experiments, instillation of naked pDNA solution onto the liver surface using a micropipette caused no significant differences in gene transfection in the applied lobe. These results suggest that it is important to select the optimal pDNA solution composition to control the gene transfection.
Hirayama
Ryu
R
Graduate School of Biomedical Sciences, Nagasaki University, Japan.
Fumoto
Shintaro
S
Nishida
Koyo
K
Nakashima
Mikiro
M
Sasaki
Hitoshi
H
Nakamura
Junzo
J
D000287:Administration, Topical; D000818:Animals; D002626:Chemistry, Pharmaceutical; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics*; D005079:Excipients; D017930:Genes, Reporter / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000502:physiology*; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology; D012996:Solutions; D014162:Transfection / Q000379:methods*
D005079:Excipients; D012996:Solutions; D012965:Sodium Chloride; D004247:DNA; D008156:Luciferases; D005947:Glucose