Sodium phosphate enhances plasmid DNA expression in vivo.
Intramuscular injection of plasmid DNA results in myofiber cell expression of proteins encoded by the DNA. The preferred vehicle for plasmid DNA injections has been saline (154 mM sodium chloride) or PBS (154 mM NaCl plus 10 mM sodium phosphate). Here, it is shown that injection of luciferase or beta-galactosidase encoding plasmid DNA in a 150 mM sodium phosphate vehicle into murine muscle resulted in a two- to seven-fold increase in transgene expression compared with DNA injected in saline or PBS. When the DNA encoded secreted alkaline phosphatase, preproinsulin or interferon, sodium phosphate vehicle increased their serum levels by two- to four-fold. When the DNA encoded mouse erythropoietin, sodium phosphate vehicle increased hematocrits by two-fold compared with DNA injected in saline. When the DNA encoded influenza nucleoprotein, sodium phosphate increased anti-nucleoprotein antibody titers by two-fold. The expression of luciferase from plasmid DNA instilled into lung was increased five-fold compared with that in vehicle without sodium phosphate. Incubation of plasmid DNA with muscle extract or serum showed that sodium phosphate protected the DNA from degradation. Thus, a change from sodium chloride to sodium phosphate vehicle can enhance the expression of plasmid DNA in a tissue, possibly by inhibiting DNA degradation. Gene Therapy (2000) 7, 1171-1182.
Department of Cell Biology, Vical Incorporated, 9373 Towne Centre Drive, San Diego, California 92121, USA.
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J
Hartikka
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Bozoukova
D
D
Jones
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R
Mahajan
M K
MK
Wloch
M
M
Sawdey
C
C
Buchner
L
L
Sukhu
K M
KM
Barnhart
A M
AM
Abai
J
J
Meek
N
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Shen
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Manthorpe
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000917:Antibody Formation; D004247:DNA / Q000276:immunology / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D004921:Erythropoietin / Q000378:metabolism; D005260:Female; D006863:Hydrogen-Ion Concentration; D007328:Insulin; D007370:Interferon Type I / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000201:enzymology; D009994:Osmolar Concentration; D010710:Phosphates / Q000494:pharmacology*; D010957:Plasmids / Q000378:metabolism*; D011384:Proinsulin / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D007328:Insulin; D007370:Interferon Type I; D010710:Phosphates; D011498:Protein Precursors; C063743:interferon omega 1; D004921:Erythropoietin; C013737:preproinsulin; D004247:DNA; D011384:Proinsulin; D008156:Luciferases; D003851:Deoxyribonucleases; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase; C018279:sodium phosphate