Correction of a Genetic Defect by Nuclear Transplantation and Combined Cell and Gene Therapy

Immune-deficient Rag2 −/− mice were used as nuclear donors for transfer into enucleated oocytes, and the resulting blastocysts were cultured to isolate an isogenic embryonic stem cell line. One of the mutated alleles in the Rag2 −/− ES cells was repaired by homologous recombination, thereby restorin...

Full description

Saved in:
Bibliographic Details
Published in:Cell 2002-04, Vol.109 (1), p.17-27
Main Authors: Rideout, William M., Hochedlinger, Konrad, Kyba, Michael, Daley, George Q., Jaenisch, Rudolf
Format: Article
Language:English
Subjects:
Animals
Blastocyst - cytology
Blastocyst - immunology
Blastocyst - metabolism
Bone Marrow Transplantation
Cell Differentiation - genetics
Cell Nucleus - genetics
Cell Nucleus - immunology
Cloning, Organism - methods
Disease Models, Animal
DNA-Binding Proteins - deficiency
DNA-Binding Proteins - genetics
Embryo Transfer
Female
Genetic Therapy
Graft Survival - genetics
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - immunology
Hematopoietic Stem Cells - metabolism
Male
Mice
Mice, Knockout
Mice, SCID
Mutation - genetics
Nuclear Transfer Techniques
Oocytes - cytology
Oocytes - immunology
Oocytes - metabolism
Polyploidy
Severe Combined Immunodeficiency - genetics
Severe Combined Immunodeficiency - immunology
Severe Combined Immunodeficiency - therapy
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Immune-deficient Rag2 −/− mice were used as nuclear donors for transfer into enucleated oocytes, and the resulting blastocysts were cultured to isolate an isogenic embryonic stem cell line. One of the mutated alleles in the Rag2 −/− ES cells was repaired by homologous recombination, thereby restoring normal Rag2 gene structure. Mutant mice were treated with the repaired ES cells in two ways. (1) Immune-competent mice were generated from the repaired ES cells by tetraploid embryo complementation and were used as bone marrow donors for transplantation. (2) Hematopoietic precursors were derived by in vitro differentiation from the repaired ES cells and engrafted into mutant mice. Mature myeloid and lymphoid cells as well as immunoglobulins became detectable 3–4 weeks after transplantation. Our results establish a paradigm for the treatment of a genetic disorder by combining therapeutic cloning with gene therapy.
ISSN:0092-8674
1097-4172
DOI:10.1016/S0092-8674(02)00681-5