Feasibility of Retroviral Vector-Mediated in utero Gene Transfer to the Fetal Rabbit

2005 ◽  
Vol 20 (6) ◽  
pp. 485-493 ◽  
Author(s):  
Rafael Moreno ◽  
Marta Rosal ◽  
Lluis Cabero ◽  
Eduard Gratacós ◽  
Josep M. Aran
Keyword(s):  
Gene Transfer ◽  
Retroviral Vector ◽  
In Utero

In utero transplantation of fetal liver cells in the mucopolysaccharidosis type VII mouse results in low-level chimerism, but overexpression of β-glucuronidase can delay onset of clinical signs

Blood ◽  
2001 ◽  
Vol 97 (6) ◽  
pp. 1625-1634 ◽  
Author(s):  
Margret L. Casal ◽  
John H. Wolfe
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Retroviral Vector ◽  
In Utero ◽  
Hematopoietic Stem ◽  
Donor Cells ◽  
Stem And Progenitor Cells ◽  
Mps Vii

Mice with the lysosomal storage disease mucopolysaccharidosis (MPS) VII, caused by a deficiency of β-glucuronidase (GUSB), have signs of disease present at birth. Bone marrow transplantation (BMT) or retroviral vector–mediated gene transfer into hematopoietic stem cells can partially correct the disease in adult mice, and BMT performed at birth results in a better clinical outcome. Thus, treatment in utero may result in further improvement. However, this must be done without cyto-ablation, and the donor cells do not have a competitive repopulating advantage over host cells. Transplantation in utero of either syngeneic fetal liver hematopoietic stem cells marked with a retroviral vector, or allogeneic donor cells that constitutively express high levels of human GUSB from a transgene, resulted in only about 0.1% engraftment in the adult. Immuno-affinity enrichment of stem and progenitor cells of 5- to 10-fold resulted in significantly higher GUSB activities at 2 months of age, but by 6 months engraftment was about 0.1%. Attempts to further increase the number of stem and progenitor cells were deleterious to the recipients. Nevertheless, GUSB expressed during the first 2 months of life in MPS VII fetuses could delay the onset of overt signs of disease. This suggests that the expression of some normal enzyme activity beginning in fetal life may offer the possibility of slowing the progression of the disease until more definitive postnatal transplantation or gene transfer to stem cells could be accomplished.

In Utero Muscle Gene Transfer

2009 ◽  
pp. 23-40
Author(s):  
Bhanu Munil Koppanati ◽  
Paula R. Clemens
Keyword(s):  
Gene Transfer ◽  
In Utero ◽  
Muscle Gene

Genes transferred by retroviral vectors into normal and mutant myoblasts in primary cultures are expressed in myotubes

1990 ◽  
Vol 10 (6) ◽  
pp. 3268-3271
Author(s):  
B F Smith ◽  
R K Hoffman ◽  
U Giger ◽  
J H Wolfe
Keyword(s):  
Gene Transfer ◽  
Primary Cultures ◽  
Muscle Cells ◽  
Retroviral Vectors ◽  
Retroviral Vector ◽  
Enzyme Deficiency ◽  
Transfer Genes

Retroviral vectors were used to transfer genes efficiently into rat and dog myoblasts in primary cultures under conditions which permitted the transduced myoblasts to differentiate into myotubes expressing the transferred genes. The transduced myotubes expressed normal markers of differentiation and were morphologically indistinguishable from uninfected myotubes. Retroviral vector-mediated gene transfer was also used to correct a genetic enzyme deficiency in mutant canine muscle cells.

scholarly journals Enhanced Green Fluorescent Protein as Selectable Marker of Retroviral-Mediated Gene Transfer in Immature Hematopoietic Bone Marrow Cells

Blood ◽  
1997 ◽  
Vol 90 (9) ◽  
pp. 3304-3315 ◽  
Author(s):  
Marti F.A. Bierhuizen ◽  
Yvonne Westerman ◽  
Trudi P. Visser ◽  
Wati Dimjati ◽  
Albertus W. Wognum ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Green Fluorescent Protein ◽  
Gene Transfer ◽  
Fluorescent Protein ◽  
Bone Marrow Cells ◽  
Retroviral Vector ◽  
Egfp Expression ◽  
Green Fluorescent ◽  
Marrow Cells

Abstract The further improvement of gene transfer into hematopoietic stem cells and their direct progeny will be greatly facilitated by markers that allow rapid detection and efficient selection of successfully transduced cells. For this purpose, a retroviral vector was designed and tested encoding a recombinant version of the Aequorea victoria green fluorescent protein that is enhanced for high-level expression in mammalian cells (EGFP). Murine cell lines (NIH 3T3, Rat2) and bone marrow cells transduced with this retroviral vector demonstrated a stable green fluorescence signal readily detectable by flow cytometry. Functional analysis of the retrovirally transduced bone marrow cells showed EGFP expression in in vitro clonogenic progenitors (GM-CFU), day 13 colony-forming unit-spleen (CFU-S), and in peripheral blood cells and marrow repopulating cells of transplanted mice. In conjunction with fluorescence-activated cell sorting (FACS) techniques EGFP expression could be used as a marker to select for greater than 95% pure populations of transduced cells and to phenotypically define the transduced cells using antibodies directed against specific cell-surface antigens. Detrimental effects of EGFP expression were not observed: fluorescence intensity appeared to be stable and hematopoietic cell growth was not impaired. The data show the feasibility of using EGFP as a convenient and rapid reporter to monitor retroviral-mediated gene transfer and expression in hematopoietic cells, to select for the genetically modified cells, and to track these cells and their progeny both in vitro and in vivo.

scholarly journals Antigen presentation in retroviral vector-mediated gene transfer in vivo

1997 ◽  
Vol 94 (5) ◽  
pp. 1943-1948 ◽  
Author(s):  
E. S. Song ◽  
V. Lee ◽  
C. D. Surh ◽  
A. Lynn ◽  
D. Brumm ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Antigen Presentation ◽  
Retroviral Vector
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