Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6461864

Title:Methods and vector constructs for making non-human animals which ubiquitously express a heterologous gene
Inventors:Soriano, Philippe; Seattle, WA, USA
Summary:This invention describes vector constructs for making transgenic nonhuman animals, such that one type has a general deletor construct while a second type has a general reporter construct. The general deletor animals express a heterologous recombinase under the control of an ubiquitously expressed endogenous promoter. Specifically, the Cre recombinase is inserted into the ROSA26 locus of the mouse. The general reporter animals have a gene flanked by sites recognized by the heterologous recombinase. This flanked sequence is operatively associated with a marker gene such that when the gene sequence flanked by sites that are recognized by the heterologous recombinase is excised, the reporter gene is expressed. Further disclosed are methods of introducing into a pluripotent cell a DNA construct comprising a heterologous DNA segment and at least 100 base pairs, homologous with a DNA sequence of a ubiquitously expressed endogenous gene locus of the pluripotent cell. In this model the DNA construct becomes integrated into the gene locus by homologous recombination, thereby inserting the heterologous DNA segment into the ubiquitously expressed endogenous gene locus such that expression of the heterologous gene segment is under the control of the promoter associated with the ubiquitously expressed endogenous gene. When the general deletor mouse is crossed with the general reporter mouse the heterologous recombinase is expressed in essentially all cells of the resultant descendants under the control of the ubiquitous promoter. Expression of the recombinase results in the excision of the desired gene in essentially all cells of the descendant animals. A conditional deletor construct and its use to derive a conditional deletor animal are also described.
Abstract:Transgenic non-human animals one having a general deletor construct and a second having a general reporter construct are described. The general deletor animals express a heterologous recombinase under the control of an ubiquitously expressed endogenous promoter. Specifically, the Cre recombinase is inserted into the ROSA26 locus of the mouse. The general reporter animals have a gene which is desired to remove flanked by sites recognized by the is heterologous recombinase. This flanked sequence is operatively associated with a marker gene such that when the gene sequence flanked by sites recognized by the heterologous recombinase is excised, the reporter gene is expressed. When the general deletor mouse is crossed with the general reporter mouse the heterologous recombinase is expressed in essentially all cells of the resultant descendants under the control of the ubiquitous promoter. Expression of the recombinase results in the excision of the desired gene in essentially all cells of the descendant animals. A conditional deletor construct and its use to derive a conditional deletor animal are also described.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6461864
Application Number:US1999000291541
Date Filed:14/04/1999
Date Published:08/10/2002
Assignee:Fred Hutchinson Cancer Research Center, Seattle, WA, USA


 
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