01. Gene therapy Boulikas.pdf - Gene therapy & Molecular Biology

Gene Therapy and Molecular Biology Vol 1, page 43
Figure 17. Endogenous mammalian β-Gal activity was not detected following incubation with X-Gal at pH>7.5 whereas bacterial β-
Gal activity was detected at pH 8.0-8.5 in airway epithelial cells in the transgenic Rosa-26 mouse. Original magnification 250X. From
Weiss DJ, Liggitt D, Clark JG (1997) In situ histochemical detection of β-galactosidase activity in lung: assessment of X-Gal reagent in
distinguishing lacZ gene expression and endogenous β-galactosidase activity. Hum Gene Ther 8, 1545-1554. With the kind permission
of the authors (Daniel Weiss, Fred Hutchinson Cancer Research Center) and Mary Ann Liebert, Inc.
Table 2. Reporter gene transfer in vivo
Gene
delivered
CAT and
luciferase
Luciferase/
CMV
construct
and lacZ
Luciferase/
CMV
lacZ and
HSV-tk/IL-
2
Human
disease
Vector Method/Goal Results Reference
none DOTM Expression of reporter genes in CMV promoter was most effective;
Zhu et al, 1993
A: mouse tissues after a single expression in vascular endothelia,
DOPE intravenous injection.
extravascular and parenchymal cells present
in lung, spleen, and heart; lower expression
in all other tissues; persisted for 9 weeks.
none Transfe To show effectiveness in gene Transient expression of luciferase in striatal Schwartz et al,
ctam transfer after intracranial injection parenchymal cells; lipospermine:DNA 1995
(DOGS of liposome-plasmid complexes charge ratio of 2 or smaller was most
:DOPE
)
into the newborn mouse.
effective in vivo.
none Cationi Distribution of luciferase gene Use of viral origins of replication and T Thierry et al,
c expression among tissues and antigen cDNA in vector sustained expression 1995
liposo persistence of expression after up to 3 months primarily in the lung in mice.
mes systemic injection.
glioma AAV Direct injection of tumors induced 30-40% of the cells along the needle track Okada et al,.
from human glioma cells into the expressed β-galactosidase; administration of 1996
brains of nude mice.
GCV to the HSV-tk/IL-2 treated animals for
6 days, resulted in a 35-fold reduction in the
mean volume of tumors compared with
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