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Stem Cell Related Patent Number US7135608
| Title: | Site-specific recombination in eukaryotes and constructs useful therefor | | Inventors: | O'Gorman, Stephen; San Diego, CA, USA
Wahl, Geoffrey; San Diego, CA, USA | | Summary: | This invention introduces novel nucleic acid constructs comprising a germline-specific promoter that is operably linked to a recombinase coding sequence. Disclosed herein are the methods by which site-specific recombinases offer a means of efficiently manipulating chromosomal sequences in mammalian cells in culture and in animals. Further described are embryonic stem cells containing recombinase nucleic acid constructs that were expressed in the male germline, and which were found to simplify current protocols for producing mice bearing homologously recombined alleles that have been secondarily rearranged by a site-specific recombinase. Recombinase coding sequences are provided, as are further applications in producing subtle, conditional or tissue-specific mutations. | | Abstract: | Site-specific recombinases provide a means of efficiently manipulating chromosomal sequences in mammalian cells in culture and in animals. Embryonic stem cells containing recombinase nucleic acid constructs that were expressed in the male germline simplify current protocols for producing mice bearing homologously recombined alleles that have been secondarily rearranged by a site-specific recombinase. In different ProCre strains, between 80% and 100% of the progeny that inherited a Cre target nucleic acid construct from males that were also heterozygous for a ProCre nucleic acid construct inherited the Cre-recombined target. ProCre nucleic acid constructs and recombined targets segregated in the first generation. When ES cells prepared from one ProCre line were transfected with vectors containing a loxP-flanked neomycin cassette, G418 resistant, homologously recombined clones, in which the loxP sites remained functional, were readily isolated. | | US Patent Website: | Click Here for Full Text of Patent | | Title Number: | US7135608 | | Application Number: | US1997000919501 | | Date Filed: | 28/08/1997 | | Date Published: | 14/11/2006 | | Assignee: | The Salk Institute for Biological Studies, La Jolla, CA, USA |
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