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Nat Biotechnol. 2003 Jun;21(6):652-9. Epub 2003 May 5.

High-throughput engineering of the mouse genome coupled with high-resolution expression analysis.

Author information

1
Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill Road, Tarrytown, New York 10591, USA.

Abstract

One of the most effective approaches for determining gene function involves engineering mice with mutations or deletions in endogenous genes of interest. Historically, this approach has been limited by the difficulty and time required to generate such mice. We describe the development of a high-throughput and largely automated process, termed VelociGene, that uses targeting vectors based on bacterial artificial chromosomes (BACs). VelociGene permits genetic alteration with nucleotide precision, is not limited by the size of desired deletions, does not depend on isogenicity or on positive-negative selection, and can precisely replace the gene of interest with a reporter that allows for high-resolution localization of target-gene expression. We describe custom genetic alterations for hundreds of genes, corresponding to about 0.5-1.0% of the entire genome. We also provide dozens of informative expression patterns involving cells in the nervous system, immune system, vasculature, skeleton, fat and other tissues.

PMID:
12730667
DOI:
10.1038/nbt822
[Indexed for MEDLINE]

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