Document Detail


Targeted gene correction of hprt mutations by 45 base single-stranded oligonucleotides.
MedLine Citation:
PMID:  12470647     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Targeted correction of a single base in a gene of an eucaryotic cell by specific oligonucleotides is a yet controversial technique. Here, we introduce the correction of point mutations in the hypoxanthine-guanine-phosphoribosyl-transferase (HPRT) gene as an additional model system to test targeted gene correction. In human, Hprt mutations cause Lesch-Nyhan syndrome. Using hamster V79 cells, we generated three cell lines with one hprt point mutation each. These cell lines were treated with specific single-stranded 45 base phosphothioate modified oligonucleotides and selected by HAT medium. The surviving clones were investigated for the correction of the respective hprt mutation. Treatment with the oligonucleotides was successful in repairing all three hprt mutations (hprt cDNA position 74, C --> T; position 151, C --> T; and position 400, G --> A). The correction efficiency was very low but reproducible. We suggest that this system allows one to investigate targeted gene correction in dependence on the target sequence and the oligonucleotides used.
Authors:
Oliver Kenner; Andrea Kneisel; Jürgen Klingler; Britta Bartelt; Günter Speit; Walther Vogel; Dieter Kaufmann
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  299     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2002 Dec 
Date Detail:
Created Date:  2002-12-09     Completed Date:  2003-01-24     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  787-92     Citation Subset:  IM    
Affiliation:
Department of Human Genetics, University of Ulm, Albert-Einstein-Allee 11, D 89070 Ulm, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Cell Line
Clone Cells
Cricetinae
DNA, Single-Stranded
Gene Targeting / methods*
Hypoxanthine Phosphoribosyltransferase / genetics*
Oligonucleotides* / chemistry
Point Mutation*
Chemical
Reg. No./Substance:
0/DNA, Single-Stranded; 0/Oligonucleotides; EC 2.4.2.8/Hypoxanthine Phosphoribosyltransferase

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


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