Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6986887

Title:Universal stem cells
Inventors:Lawman, Patricia; Chipley, FL, USA
Lawman, Michael J. P.; Chipley, FL, USA
Summary:Herein are described methods of production of stem cells expressing a heterologous histocompatibility allele, via the deletion of regions of the histocompatibility gene complex and replacement of these regions with a polynucleotide encoding a heterologous allele. Described herein are methods and materials pertaining to the preparation of multipotent stem cells with a preselected expression of MHC antigens, including the use of targeting vectors that are capable of gene knockout, insertion of site-specific recombination cassettes, and the replacement of histocompatibility alleles in the stem cell. Further disclosed are novel knockout vectors that are used to delete designated regions of a chromosome, after which recombination cassette vectors are then used to delete the same region on a second chromosome and to deposit a site-specific recombination cassette which can be utilized by replacement vectors for inserting the new MHC genes on the chromosome of the engineered cell. Methods are described by which the site-specific recombination is catalyzed such that the transgene becomes incorporated into the second chromosome, thereby providing a stem cell with altered histocompatibility phenotype. Properties of related cells, tissues, and transgenic mammals are provided, as are therapeutic applications in the generation of histocompatible tissues and organs for transplantation.
Abstract:The subject invention pertains to materials and methods for preparing multi-potential stem cells having a pre-selected expression of MHC antigens. Stem cells of the subject invention can be used to generate histocompatible tissues/organs for transplantation. The process of the subject invention comprises the use of targeting vectors capable of gene knockout, insertion of site-specific recombination cassettes, and the replacement of histocompatibility alleles in the stem cell. Novel knockout vectors are used to delete designated regions of one chromosome. Recombination cassette vectors are then used to delete the same region on the second chromosome and deposit a site-specific recombination cassette which can be utilized by replacement vectors for inserting the new MHC genes on the chromosome of the engineered cell. The subject invention also pertains to cells, tissues, and transgenic mammal prepared using the methods and materials of the invention.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6986887
Application Number:US2002000186231
Date Filed:28/06/2002
Date Published:17/01/2006
Assignee:Morphogenesis, Inc., Oldsmar, FL, USA


 
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