Generation of large genomic DNA deletions

Details Generation of large genomic DNA deletions Generation of large genomic DNA deletions

1997-04-16

1999-12-07

McKelvey, Terry

Chemistry: molecular biology and microbiology

Process of mutation, cell fusion, or genetic modification

Introduction of a polynucleotide molecule into or...

435325, C12N 1500, C12N 510

Patent

active

059982098

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.

REFERENCES:
Zhang et al, Molecular and Cellular Biology vol. 14(4): pp. 2404-2410 (Apr. 1994).
Ramirez-Solis et al, Nature, vol. 378: pp. 720-724 (Dec. 14, 1995).

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