Document Detail

High-throughput engineering of the mouse genome coupled with high-resolution expression analysis.
MedLine Citation:
PMID:  12730667     Owner:  NLM     Status:  MEDLINE    
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.
David M Valenzuela; Andrew J Murphy; David Frendewey; Nicholas W Gale; Aris N Economides; Wojtek Auerbach; William T Poueymirou; Niels C Adams; Jose Rojas; Jason Yasenchak; Rostislav Chernomorsky; Marylene Boucher; Andrea L Elsasser; Lakeisha Esau; Jenny Zheng; Jennifer A Griffiths; Xiaorong Wang; Hong Su; Yingzi Xue; Melissa G Dominguez; Irene Noguera; Richard Torres; Lynn E Macdonald; A Francis Stewart; Thomas M DeChiara; George D Yancopoulos
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Publication Detail:
Type:  Evaluation Studies; Journal Article     Date:  2003-05-05
Journal Detail:
Title:  Nature biotechnology     Volume:  21     ISSN:  1087-0156     ISO Abbreviation:  Nat. Biotechnol.     Publication Date:  2003 Jun 
Date Detail:
Created Date:  2003-05-30     Completed Date:  2003-08-15     Revised Date:  2006-04-06    
Medline Journal Info:
Nlm Unique ID:  9604648     Medline TA:  Nat Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  652-9     Citation Subset:  IM    
Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill Road, Tarrytown, New York 10591, USA.
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MeSH Terms
Animals, Genetically Modified / genetics,  metabolism
Chromosomes, Artificial, Bacterial / genetics*,  metabolism*
Electroporation / methods
Gene Expression Profiling / methods*
Gene Targeting / methods
Genetic Engineering / methods*
Mice / genetics
Mutagenesis, Insertional / methods
Mutagenesis, Site-Directed
Quality Control
Recombinant Proteins / genetics,  metabolism
Stem Cells / metabolism
Reg. No./Substance:
0/Recombinant Proteins
Comment In:
Nat Biotechnol. 2003 Jun;21(6):625-7   [PMID:  12776146 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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