Secreted Human Placental Alkaline Phosphatase as a Reporter Gene for in Vivo Gene Transfer

1999 ◽  
Vol 271 (2) ◽  
pp. 187-189 ◽  
Author(s):  
Mickaël Bettan ◽  
Raphaël Darteil ◽  
Daniel Scherman
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Transfer ◽  
Reporter Gene ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase ◽  
In Vivo Gene Transfer

scholarly journals The Promoter of the Rat Gonadotropin-Releasing Hormone Receptor Gene Directs the Expression of the Human Placental Alkaline Phosphatase Reporter Gene in Gonadotrope Cells in the Anterior Pituitary Gland as well as in Multiple Extrapituitary Tissues

Endocrinology ◽  
2004 ◽  
Vol 145 (2) ◽  
pp. 983-993 ◽  
Author(s):  
Anne Granger ◽  
Valérie Ngô-Muller ◽  
Christian Bleux ◽  
Céline Guigon ◽  
Hanna Pincas ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Reporter Gene ◽  
Transgene Expression ◽  
Pituitary Cells ◽  
Placental Alkaline Phosphatase ◽  
R Gene ◽  
Glycoprotein Hormone ◽  
Steroidogenic Factor 1 ◽  
Functional Sites ◽  
Human Placental Alkaline Phosphatase

Abstract Previous studies dealing with the mechanisms underlying the tissue-specific and regulated expression of the GnRH receptor (GnRH-R) gene led us to define several cis-acting regulatory sequences in the rat GnRH-R gene promoter. These include functional sites for steroidogenic factor 1, activator protein 1, and motifs related to GATA and LIM homeodomain response elements as demonstrated primarily in transient transfection assays in mouse gonadotrope-derived cell lines. To understand these mechanisms in more depth, we generated transgenic mice bearing the 3.3-kb rat GnRH-R promoter linked to the human placental alkaline phosphatase reporter gene. Here we show that the rat GnRH-R promoter drives the expression of the reporter gene in pituitary cells expressing the LHβ and/or FSHβ subunit but not in TSHβ- or GH-positive cells. Furthermore, the spatial and temporal pattern of the transgene expression during the development of the pituitary was compatible with that characterizing the emergence of the gonadotrope lineage. In particular, transgene expression is colocalized with the expression of the glycoprotein hormone α-subunit at embryonic day 13.5 and with that of steroidogenic factor 1 at later stages of pituitary development. Transgene expression was also found in specific brain areas, such as the lateral septum and the hippocampus. A single promoter is thus capable of directing transcription in highly diverse tissues, raising the question of the different combinations of transcription factors that lead to such a multiple, but nevertheless cell-specific, expressions of the GnRH-R gene.

scholarly journals Replication-competent retroviral vectors encoding alkaline phosphatase reveal spatial restriction of viral gene expression/transduction in the chick embryo.

1993 ◽  
Vol 13 (4) ◽  
pp. 2604-2613 ◽  
Author(s):  
D M Fekete ◽  
C L Cepko
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Transfer ◽  
Time Window ◽  
Expression Patterns ◽  
Viral Gene Expression ◽  
Retroviral Vectors ◽  
Viral Gene ◽  
Neural Cells ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase

Replication-competent avian retroviruses, capable of transducing and expressing up to 2 kb of nonviral sequences, are now available to effect widespread gene transfer in chicken (chick) embryos (S. H. Hughes, J. J. Greenhouse, C. J. Petropoulos, and P. Sutrave, J. Virol. 61:3004-3012, 1987). We have constructed novel avian retroviral vectors that encode human placental alkaline phosphatase as a marker whose expression can be histochemically monitored. These vectors have been tested for expression by introducing them into the embryonic chick nervous system. They have revealed that the expression of retrovirally transduced genes can be spatially and temporally limited without the need for tissue-specific promoters. By varying the site and time of infection, targeted gene transfer can be confined to selected populations of neural cells over the course of several days, a time window that is sufficient for many key developmental processes. The capability of differentially infecting specific target populations may avoid confounding variables such as detrimental effects of a transduced gene on processes unrelated to the cells or tissue of interest. These vectors and methods thus should be useful in studies of the effect of transduced genes on the development of various organs and tissues during avian embryogenesis. In addition, the vectors will facilitate studies aimed at an understanding of viral infection and expression patterns.

Phenotypic and immunologic factors affecting plasmid-mediated in vivo gene transfer to rat diaphragm

1996 ◽  
Vol 270 (6) ◽  
pp. L1023-L1030 ◽  
Author(s):  
B. J. Petrof ◽  
G. Acsadi ◽  
J. Bourdon ◽  
N. Matusiewicz ◽  
L. Yang
Keyword(s):  
Gene Expression ◽  
Gene Transfer ◽  
Reporter Gene ◽  
Transfection Efficiency ◽  
Foreign Gene ◽  
Rat Diaphragm ◽  
Foreign Gene Expression ◽  
Hindlimb Muscles ◽  
In Vivo Gene Transfer

Little is known about the molecular mechanisms governing adaptive responses of the diaphragm in the setting of lung disease. By permitting the study of regulatory elements and the effects of overexpressing genes of interest, direct in vivo gene transfer to the diaphragm could be used as a tool to address such questions. Therefore, we evaluated parameters affecting transfection efficiency and duration of foreign gene expression in the diaphragm after plasmid-mediated gene transfer. Reporter gene constructs were injected into adult rat diaphragm and hindlimb muscles. Transfection efficiency at 8-10 days postinjection was decreased in large caliber ( > 1,000 microns2) and type II myosin heavy chain (MHC)-expressing fibers. There were also strong trends toward augmented transfection efficiency in type I MHC- and embryonic MHC-expressing fibers. All diaphragms demonstrated evidence of muscle injury and inflammatory cell infiltrates at this early time point. By 30 days postinjection, however, neither inflammation nor reporter gene expression was detectable in diaphragm or hindlimb muscles of immunocompetent animals. By contrast, immunosuppressed rats (given cyclosporine; 15 mg.kg-1. day-1) showed high levels of foreign gene expression at 30 days postinjection, which remained stable up to 60 days. Therefore, exploitation of plasmid-mediated in vivo gene transfer as a tool for studying regulated gene expression in the diaphragm may be facilitated by the use of immunodeficient animal models.

Replication-competent retroviral vectors encoding alkaline phosphatase reveal spatial restriction of viral gene expression/transduction in the chick embryo

1993 ◽  
Vol 13 (4) ◽  
pp. 2604-2613
Author(s):  
D M Fekete ◽  
C L Cepko
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Transfer ◽  
Time Window ◽  
Expression Patterns ◽  
Viral Gene Expression ◽  
Retroviral Vectors ◽  
Viral Gene ◽  
Neural Cells ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase

Replication-competent avian retroviruses, capable of transducing and expressing up to 2 kb of nonviral sequences, are now available to effect widespread gene transfer in chicken (chick) embryos (S. H. Hughes, J. J. Greenhouse, C. J. Petropoulos, and P. Sutrave, J. Virol. 61:3004-3012, 1987). We have constructed novel avian retroviral vectors that encode human placental alkaline phosphatase as a marker whose expression can be histochemically monitored. These vectors have been tested for expression by introducing them into the embryonic chick nervous system. They have revealed that the expression of retrovirally transduced genes can be spatially and temporally limited without the need for tissue-specific promoters. By varying the site and time of infection, targeted gene transfer can be confined to selected populations of neural cells over the course of several days, a time window that is sufficient for many key developmental processes. The capability of differentially infecting specific target populations may avoid confounding variables such as detrimental effects of a transduced gene on processes unrelated to the cells or tissue of interest. These vectors and methods thus should be useful in studies of the effect of transduced genes on the development of various organs and tissues during avian embryogenesis. In addition, the vectors will facilitate studies aimed at an understanding of viral infection and expression patterns.

1860: Generation of Fluorescent Sperm by Use of in VIVO Gene Transfer to Hamster Testes

2007 ◽  
Vol 177 (4S) ◽  
pp. 617-618
Author(s):  
Hiraki Kubota ◽  
Kevin Coward ◽  
Olivia Hibbitt ◽  
Nilendran Prathalingam ◽  
William Holt ◽  
...  
Keyword(s):  
Gene Transfer ◽  
In Vivo Gene Transfer
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