Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5837536

Title:Expression of human multidrug resistance genes and improved selection of cells transduced with such genes
Inventors:McDonagh, Kevin T.; Silver Spring, MD, USA
Nienhuis, Arthur; Bethesda, MD, USA
Tolstoshev, Paul; Potomac, MD. USA
Summary:Described herein are isolated DNA or RNA sequences for the human multi-drug-resistant gene MDR1, which encodes p-glycoprotein such that at least one base in a splice region of the DNA encoding the p-glycoprotein is changed. Claims of the invention include prevention of a truncated p-glycoprotein upon expression as a result of such a mutation. Further disclosed are methods of identifying cells which express the human MDR1 gene in a cell population that has been transduced with an expression vehicle including a human MDR1 gene, via a process of contacting the cell population with a staining material such as rhodamine 123 and identifying cells which express the human MDR1 gene based on differentiation in color among the cells of the cell population. Therapeutic applications include gene therapy, such as in the identification of retroviral producer clones which facilitate MDR1 gene transfer into primary cells, and the repopulating of hematopoietic stem cells that have been genetically engineered with the human MDR1 gene for multi-drug resistance in cancer therapy.
Abstract:A DNA sequence for a human mdr1 gene, which encodes p-glycoprotein, wherein at least one base in a splice region of the DNA encoding p-glycoprotein is changed. Such a mutation prevents truncation of the p-glycoprotein upon expression thereof. There is also provided a method of identifying cells which express the human mdr1 gene in a cell population that has been transduced with an expression vehicle including a human mdr1 gene. The method comprises contacting the cell population with a staining material, such as rhodamine 123, and identifying cells which express the human mdr1 gene based on differentiation in color among the cells of the cell population. This method has allowed identification of retroviral producer clones facilitate mdr gene transfer into primary cells. Repopulating hematopoietic stem cells have been genetically engineered with the human mdr1 gene.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5837536
Application Number:US1996000583276
Date Filed:05/01/1996
Date Published:17/11/1998
Assignee:Genetic Therapy, Inc., Gaithersburg, MD, USA The USA as represented by the Department of Health and Human Services, Washington, DC, USA


 
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