Methods for genetic diversification in gene conversion active cells
AssigneeGSF-Forschungszentrum fur Umwelt und Gesundheit GmbH
InventorsJean-Marie Buerstedde, Hiroshi Arakawa
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Abstract
The invention relates to a modified lymphoid cell having gene conversion fully or partially replaced by hypermutation, wherein said cell has no deleterious mutations in genes encoding paralogues and analogues of the RAD51 protein, and wherein said cell is capable of directed and selective genetic diversification of a target nucleic acid by hypermutation or a combination of hypermutation and gene conversion. The invention also relates to a method for diversifying any transgenic target gene in said cell. Preferably, the target gene is integrated into the immunoglobulin light or heavy chain locus by targeted integration.
Claims43
A genetically modified lymphoid cell having gene conversion fully or partially replaced by hypermutation, wherein said cell has no deleterious mutations in genes encoding paralogues and analogues of the RAD5 1 protein.
A non-human transgenic animal containing a lymphoid cell having gene conversion fully or partially replaced by hypermutation, wherein said cell has no deleterious mutations in genes encoding paralogues and analogues of the RAD51 protein, and wherein said cell is capable of directed and selective genetic diversification of a transgenic target nucleic acid by hypermutation or a combination of hypermutation and gene conversion.
A method for preparing a cell capable of directed and selective genetic diversification of a target nucleic acid by hypermutation or a combination of hypermutation and gene conversion comprising (a) transfecting a lymphoid cell capable of gene conversion with a genetic construct containing the target nucleic acid, and (b) identifying a cell having the endogenous V-gene or a fragment thereof replaced with the target nucleic acid.
Dependencies are inferred from the claim text.