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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 homologous recombination for a locus in a villin gene via the introduction of an I-SceI restriction site. The invention relates to a method of constructing a villin gene hybrid by inserting an I-Sce I restriction site next to or within a gene or cDNA encoding a villin protein, such that the location of insertion of the I-Sce I restriction site is chosen to provide a first downstream part and a second upstream part from the site, which contains at least twelve nucleotides of the gene or cDNA encoding the villin protein. Claims of the invention include a high frequency of homologous recombination events that result from the insertion of the restriction site. Therapeutic applications of the villin gene hybrid include its use in the transfection of eukaryotic cells such as embryonic stem cells. |
| Abstract: | This invention relates to a method of constructing a villin gene hybrid by inserting an I-Sce I restriction site next to or within a gene or cDNA encoding a villin protein. The insertion site of the I-Sce I restriction site is chosen as to provide a first downstream part and a second upstream part from the site, containing at least twelve nucleotides of the gene or cDNA encoding the villin protein. Furthermore, the insertion of the restriction permits a high frequency of homologous recombination events. The villin gene hybrid may be used to transfect eukaryotic cells, and particularly, embryonic stem cells. |
| US Patent Website: | Click Here for Full Text of Patent |
| Title Number: | US5830729 |
| Application Number: | US1996000693948 |
| Date Filed: | 07/08/1996 |
| Date Published: | 03/11/1998 |
| Assignee: | Institut Pasteur, Paris, France Institut Curie, Paris, France Centre National de la Recherche Scientifique, Paris, France |