DNA Oligonucleotides and Plasmids Perform Equally as Donors for Targeted Gene Conversion
Identifieur interne : 000C11 ( Istex/Corpus ); précédent : 000C10; suivant : 000C12DNA Oligonucleotides and Plasmids Perform Equally as Donors for Targeted Gene Conversion
Auteurs : Stefan Wagner ; Judi Mccracken ; Sally Cole ; Götz LaibleSource :
- 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
Links to Exploration step
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<front><div type="abstract" xml:lang="en">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.</div>
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<abstract lang="en">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.</abstract>
<subject lang="en"><genre>Keywords</genre>
<topic>Gene conversion</topic>
<topic>Green fluorescent protein</topic>
<topic>Plasmid</topic>
<topic>Oligonucleotide</topic>
<topic>Transfection</topic>
</subject>
<relatedItem type="host"><titleInfo><title>Biochemical Genetics</title>
</titleInfo>
<titleInfo type="abbreviated"><title>Biochem Genet</title>
</titleInfo>
<genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre>
<originInfo><publisher>Springer</publisher>
<dateIssued encoding="w3cdtf">2010-11-17</dateIssued>
<copyrightDate encoding="w3cdtf">2010</copyrightDate>
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<subject><genre>Biomedicine</genre>
<topic>Medical Microbiology</topic>
<topic>Zoology</topic>
<topic>Biochemistry, general</topic>
<topic>Human Genetics</topic>
</subject>
<identifier type="ISSN">0006-2928</identifier>
<identifier type="eISSN">1573-4927</identifier>
<identifier type="JournalID">10528</identifier>
<identifier type="IssueArticleCount">12</identifier>
<identifier type="VolumeIssueCount">12</identifier>
<part><date>2010</date>
<detail type="volume"><number>48</number>
<caption>vol.</caption>
</detail>
<detail type="issue"><number>11-12</number>
<caption>no.</caption>
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<extent unit="pages"><start>897</start>
<end>908</end>
</extent>
</part>
<recordInfo><recordOrigin>Springer Science+Business Media, LLC, 2010</recordOrigin>
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<identifier type="ark">ark:/67375/VQC-X6FX9VZK-Z</identifier>
<identifier type="DOI">10.1007/s10528-010-9370-z</identifier>
<identifier type="ArticleID">9370</identifier>
<identifier type="ArticleID">s10528-010-9370-z</identifier>
<accessCondition type="use and reproduction" contentType="copyright">Springer Science+Business Media, LLC, 2010</accessCondition>
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<recordOrigin>Springer Science+Business Media, LLC, 2010</recordOrigin>
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