Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6638768

Title:Procedure for specific replacement of a copy of a gene present in the recipient genome by the integration of a gene different from that where the integration is made
Inventors:Le Mouellic, Herve; Paris, France
Brulet, Philippe; Maurepas, France
Summary:This invention describes the specific replacement of a gene by targeted insertion of a heterologous gene, carried on a vector having homologous flanking sequences which reconstruct the complete gene. The method relates to a process for targeted replacement of at least a part of an endogenous gene by at least a part of a foreign gene, or targeted insertion of at least a part of a foreign gene at a targeted site of an endogenous gene in a cell by homologous recombination. Further described are first and second flanking DNA sequences homologous to their respective genomic sequences situated on one side or the other of the targeted site of the endogenous gene that is to be replaced. Also provided are methods of cellular selection and transfection with the vector, and specific properties of the vector.
Abstract:The present invention relates to a process for targeted replacement of at least a part of an endogenous gene by at least a part of a foreign gene or targeted insertion of at least a part of a foreign gene at a targeted site in an endogenous gene in a cell by homologous recombination. This process includes (A) providing a vector which contains (1) at least a part of the foreign gene which is heterologous with respect to the endogenous gene, (2) a first flanking DNA sequence homologous to a first genomic sequence situated on one side of the part of the endogenous gene to be replaced or the targeted site; and (3) a second flanking DNA sequence homologous to a second genomic sequence situated on the other side of the part of the endogenous gene to be replaced or the targeted site, the foreign gene being located between the first and second flanking DNA sequences and is complementary to the part of the endogenous gene to be replaced; (B) transfecting a cell with the vector; (C) and selecting a transfected cell that contains the foreign gene at the targeted site or where the part of the endogenous gene is to be replaced. This invention also pertains to the vector, and the cell transfected by the vector.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6638768
Application Number:US1995000466699
Date Filed:06/06/1995
Date Published:28/10/2003
Assignee:Institut Pasteur, Paris, France


 
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