DNA Oligonucleotides and Plasmids Perform Equally as Donors for Targeted Gene Conversion
Identifieur interne : 002731 ( Main/Exploration ); précédent : 002730; suivant : 002732DNA Oligonucleotides and Plasmids Perform Equally as Donors for Targeted Gene Conversion
Auteurs : Stefan Wagner [Nouvelle-Zélande] ; Judi Mccracken [Nouvelle-Zélande] ; Sally Cole [Nouvelle-Zélande] ; Götz Laible [Nouvelle-Zélande]Source :
- Biochemical Genetics [ 0006-2928 ] ; 2010-12-01.
English descriptors
Abstract
Abstract: Site-specific gene modifications in cells are initiated by the introduction of exogenous DNA. We used a recently established cell assay to compare the ability of DNA donors to induce a single point mutation that converts a target gene encoding blue fluorescent protein (BFP) into expressing green fluorescent protein (GFP). In a chromosomal assay with cells stably expressing BFP, we showed that fluorescently labeled single-stranded oligonucleotides and a donor plasmid cotranscribing a red fluorescent protein provide similar efficiencies in triggering BFP–GFP conversions. In transient cotransfections, an isogenic donor plasmid comprising a nonfunctional GFP gene yielded a greater efficiency for the conversion of the BFP target gene than a nonisogenic donor, and all plasmid donors were superior to oligonucleotides.
Url:
DOI: 10.1007/s10528-010-9370-z
Affiliations:
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We used a recently established cell assay to compare the ability of DNA donors to induce a single point mutation that converts a target gene encoding blue fluorescent protein (BFP) into expressing green fluorescent protein (GFP). In a chromosomal assay with cells stably expressing BFP, we showed that fluorescently labeled single-stranded oligonucleotides and a donor plasmid cotranscribing a red fluorescent protein provide similar efficiencies in triggering BFP–GFP conversions. In transient cotransfections, an isogenic donor plasmid comprising a nonfunctional GFP gene yielded a greater efficiency for the conversion of the BFP target gene than a nonisogenic donor, and all plasmid donors were superior to oligonucleotides.</div></front></TEI><affiliations><list><country><li>Nouvelle-Zélande</li></country></list><tree><country name="Nouvelle-Zélande"><noRegion><name sortKey="Wagner, Stefan" sort="Wagner, Stefan" uniqKey="Wagner S" first="Stefan" last="Wagner">Stefan Wagner</name></noRegion><name sortKey="Cole, Sally" sort="Cole, Sally" uniqKey="Cole S" first="Sally" last="Cole">Sally Cole</name><name sortKey="Laible, Gotz" sort="Laible, Gotz" uniqKey="Laible G" first="Götz" last="Laible">Götz Laible</name><name sortKey="Mccracken, Judi" sort="Mccracken, Judi" uniqKey="Mccracken J" first="Judi" last="Mccracken">Judi Mccracken</name><name sortKey="Wagner, Stefan" sort="Wagner, Stefan" uniqKey="Wagner S" first="Stefan" last="Wagner">Stefan Wagner</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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