Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6534314

Title:Methods and compositions for transforming cells
Inventors:Bouhassira, Eric; Hastings on Hudson, NY, USA
Summary:Described herein are methods and compositions for site-specific recombination in a selected cell via the use of acceptor and donor vectors containing incompatible lox sites. The invention relates to a process of transforming cells which results in the efficient and stable site-specific integration of transgenes, such that transformation is achieved by providing an acceptor DNA containing two inverted lox sequences and a donor DNA containing the same two inverted lox sequences, which is followed by contacting the acceptor and donor DNA with a recombinase such as Cre or Flp which causes recombination at the lox sequences contained in the DNAs. Further disclosed are methods by which the acceptor DNA is preferably integrated into the genome of a cell, such as an embryonic stem cell or a fertilized egg, prior to recombination, such that the acceptor DNA may optionally contain a negatively selectable marker to allow for screening of cells which have undergone the desired site-specific recombination such as DNA cassette exchange. Therapeutic applications in gene therapy are included.
Abstract:Methods and compositions for transforming cells, resulting in efficient and stable site-specific integration of transgenes, are disclosed. Transformation is achieved by providing an acceptor DNA containing two inverted lox sequences and a donor DNA containing the same two inverted lox sequences, and then contacting the acceptor and donor DNA with a recombinase (e.g., Cre or Flp) which causes recombination at the lox sequences contained in the DNAs. Prior to recombination, the acceptor DNA is preferably integrated into the genome of a cell, such as an embryonic stem cell or a fertilized egg. The acceptor DNA optionally may further contain a negatively selectable marker to allow for screening of cells which have undergone the desired site-specific recombination (e.g., DNA cassette exchange).
US Patent Website:Click Here for Full Text of Patent
Title Number:US6534314
Application Number:US1999000293303
Date Filed:16/04/1999
Date Published:18/03/2003
Assignee:Massachusetts Institute of Technology, Cambridge, MA, USA


 
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