Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5849991

Title:Mice homozygous for an inactivated .alpha. 1,3-galactosyl transferase gene
Inventors:d'Apice, Anthony J. F.; Balwyn, Australia
Pearse, Martin J.; Mordialloc, Australia
Robins, Allan J.; Waterloo Corner, Australia
Crawford, Robert J.; West Lake Shores, Australia
Rathjen, Peter D.; Blackwood, Australia
Summary:Described herein are a novel alpha-1,3-galactosyltransferase and a leukemia inhibiting factor, methods and uses thereof. The invention relates to a novel DNA construct comprising a disrupted mouse ?a-1,3 galactosyltransferase gene, wherein the disruption is due to the insertion of an exogenous sequence into the gene such that the disruption prevents expression of functional ?a-1,3 galactosyltransferase. Further disclosed are properties of human pre-formed xenoantibodies and their role in the hyperacute rejection response in human xenotransplantation. Methods and materials are provided for removing or neutralizing such antibodies, as is a process for the reduction and elimination of epitopes in the donor organs that are recognized by the antibodies. Also provided is a process for the formation of such epitopes from the enzyme alpha -1,3 galactosyltransferase, as are properties of the porcine gene encoding alpha -1,3 galactosyltransferase. Claims of the invention include methods and materials for inactivating the alpha -1,3 galactosyltransferase gene in mammalian cells, embryos and target cells, thereby generating knock out mice. Further claimed is a novel leukemia inhibitory factor (T-LIF) with therapeutic and experimental applications in the maintenance of embryonic stem cells and primordial germ cells in culture, for the inactivation of the transferase gene, and for the elimination of hyperacute regions in human transplants.
Abstract:Human pre-formed xenoantibodies play an important role in the hyperacute rejection response in human xenotransplantation. Disclosed are materials and methods for removing or neutralizing such antibodies. Also disclosed are materials and methods for reducing or eliminating the epitopes in the donor organs that are recognized by such antibodies. Such epitopes are formed as the result of activity by the enzyme .alpha.-1,3 galactosyltransferase. The porcine gene encoding .alpha.-1,3 galactosyltransferase is disclosed, as are materials and methods for inactivating (knocking out) the .alpha.-1,3 galactosyltransferase gene in mammalian cells and embryos. Included are nucleic acid constructs useful for inactivating the .alpha.-1,3 galactosyltransferase gene in a target cell. Also disclosed is a novel leukemia inhibitory factor (T-LIF) that is useful for maintenance of embryonic stem cells and primordial germ cells in culture.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5849991
Application Number:US1995000378617
Date Filed:26/01/1995
Date Published:15/12/1998
Assignee:Bresatch Limited, Adelaide, Australia St. Vincent's Hospital, Victoria, Australia


 
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