Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6566579

Title:I-Sce I induced gene replacement and gene conversion in embryonic stem cells
Inventors:Jaisser, Frederic; Malakoff, France
Cohen-Tannoudji, Michel; Paris, France
Robine, Sylvie; Vanves, France
Choulika, Andre; Paris, France
Louvard, Daniel; Sceaux, France
Babinet, Charles; Paris, France
Summary:Described herein is a method of introducing a deoxyribonucleic acid sequence into mouse embryonic stem cell for gene replacement in ovo. The invention relates to methods by which a heterologous DNA sequence is introduced into a mouse embryonic stem cell wherein the DNA sequence is inserted by homologous recombination into a villin gene/I-SceI hybrid by creating a double strand break with I-SceI meganuclease. The mouse embryonic stem cells are subsequently used to generate a transgenic mouse comprising the heterologous DNA sequence. Further disclosed are methods by which such techniques may be used for gene replacement in ovo wherein a mouse oocyte containing a villin gene/I-SceI hybrid within its genome exists or is first generated.
Abstract:The invention relates to methods of introducing a heterologous DNA sequence into a mouse embryonic stem cell wherein the DNA sequence is inserted by homologous recombination into a villin gene/I-SceI hybrid by creating a double strand break with I-SceI meganuclease. Subsequently, the mouse embryonic stem cells can be used to generate a transgenic mouse comprising the heterologous DNA sequence. Additionally, the methods can be used for gene replacement in ovo where a mouse oocyte containing a villin gene/I-SceI hybrid within its genome exists or is first generated. More generally, the methods can be used for the targeted insertion of a heterologous DNA sequence into any cell containing a villin gene/I-SceI hybrid sequence within its genome.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6566579
Application Number:US1998000116834
Date Filed:17/07/1998
Date Published:20/05/2003
Assignee:Institut Pasteur, Paris, France Institut Curie, Paris, France Centre Nationale de la Recherche Scientifique, Paris, France


 
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