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| 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 |