Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6884622

Title:Method for preparing a mammalian cell deficient in HPRT
Inventors:Jokobovits, Aya; Menlo Park, CA, USA
Summary:This invention describes a method by which mammalian cells are deficient in HPRT by genomic deletion. The method involves modification of the genome of the cell containing the wild-type locus via introduction of a construct comprising two regions of sequences which are homologous to 5' and 3' flanking the sequences of the targeted regions to be deleted in the locus. Further described are uses of a replacement-type targeting construct to delete large fragments of genomic DNA by such gene targeting. After transfection of the targeting construct into the desired cell line, gene targeted-mediated deletions are identified by selection and further characterized. Uses are recommended for various cases involving large genomic deletion. Examples of suitable loci are given to include MHC Class I and II antigens and immunoglobulin genes, such as the variable and constant regions of kappa, lambda, or heavy chains.
Abstract:The method of the invention provides the use of a replacement-type targeting construct to delete large fragments of genomic DNA by gene targeting. The replacement targeting construct, which may contain a selectable marker, is constructed to contain two regions of sequences which are homologous to the 5′ and 3′ flanking sequences of the targeted locus. After transfection of the targeting construct into the desired cell line, gene targeted-mediated deletions are identified by selection and further characterized. The invention is useful in any situation where one would want to create a large genomic deletion. Examples of suitable loci include MHC Class I and II antigens and immunoglobulin genes, including, for example, variable and constant region of kappa, lambda, or heavy chains.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6884622
Application Number:US2000000718717
Date Filed:22/11/2000
Date Published:26/04/2005
Assignee:Abgenix, Inc., Fremont, CA, USA


 
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