Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2
Identifieur interne : 006177 ( Main/Exploration ); précédent : 006176; suivant : 006178Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2
Auteurs : Tú Nguyen-Dumont [France] ; Lars P. Jordheim [France] ; Jocelyne Michelon [France] ; Nathalie Forey [France] ; Sandrine Mckay-Chopin [France] ; Olga Sinilnikova [France] ; Florence Le Calvez-Kelm [France] ; Melissa C. Southey [Australie] ; Sean V. Tavtigian [États-Unis] ; Fabienne Lesueur [France]Source :
- BMC Medical Genomics [ 1755-8794 ] ; 2011.
Abstract
The gene
We implemented an assay based on high-resolution melting (HRM) curve analysis and developed an analysis tool for DAE assessment.
We observed allelic expression imbalance in 4 of the 41 LCLs examined. All four were carriers of the truncating mutation 1100delC. We confirmed previous findings that this mutation induces non-sense mediated mRNA decay. In our series, we ruled out the possibility of a functional sequence variant located in the promoter region or in a regulatory element of
Our results support that HRM is a sensitive and accurate method for DAE assessment. This approach would be of great interest for high-throughput mutation screening projects aiming to identify genes carrying functional regulatory polymorphisms.
Url:
DOI: 10.1186/1755-8794-4-39
PubMed: 21569354
PubMed Central: 3112061
Affiliations:
- Australie, France, États-Unis
- Auvergne-Rhône-Alpes, Rhône-Alpes, Utah, Victoria (État)
- Lyon, Melbourne
- Université de Melbourne
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Le document en format XML
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<author><name sortKey="Lesueur, Fabienne" sort="Lesueur, Fabienne" uniqKey="Lesueur F" first="Fabienne" last="Lesueur">Fabienne Lesueur</name>
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<country xml:lang="fr">France</country>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>The gene <italic>CHEK2 </italic>
encodes a checkpoint kinase playing a key role in the DNA damage pathway. Though <italic>CHEK2 </italic>
has been identified as an intermediate breast cancer susceptibility gene, only a small proportion of high-risk families have been explained by genetic variants located in its coding region. Alteration in gene expression regulation provides a potential mechanism for generating disease susceptibility. The detection of differential allelic expression (DAE) represents a sensitive assay to direct the search for a functional sequence variant within the transcriptional regulatory elements of a candidate gene. We aimed to assess whether <italic>CHEK2 </italic>
was subject to DAE in lymphoblastoid cell lines (LCLs) from high-risk breast cancer patients for whom no mutation in <italic>BRCA1</italic>
or <italic>BRCA2</italic>
had been identified.</p>
</sec>
<sec><title>Methods</title>
<p>We implemented an assay based on high-resolution melting (HRM) curve analysis and developed an analysis tool for DAE assessment.</p>
</sec>
<sec><title>Results</title>
<p>We observed allelic expression imbalance in 4 of the 41 LCLs examined. All four were carriers of the truncating mutation 1100delC. We confirmed previous findings that this mutation induces non-sense mediated mRNA decay. In our series, we ruled out the possibility of a functional sequence variant located in the promoter region or in a regulatory element of <italic>CHEK2 </italic>
that would lead to DAE in the transcriptional regulatory milieu of freely proliferating LCLs.</p>
</sec>
<sec><title>Conclusions</title>
<p>Our results support that HRM is a sensitive and accurate method for DAE assessment. This approach would be of great interest for high-throughput mutation screening projects aiming to identify genes carrying functional regulatory polymorphisms.</p>
</sec>
</div>
</front>
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</author>
<author><name sortKey="Gurd, S" uniqKey="Gurd S">S Gurd</name>
</author>
<author><name sortKey="Gaudin, T" uniqKey="Gaudin T">T Gaudin</name>
</author>
<author><name sortKey="Dore, C" uniqKey="Dore C">C Dore</name>
</author>
<author><name sortKey="Lemire, M" uniqKey="Lemire M">M Lemire</name>
</author>
<author><name sortKey="Lepage, P" uniqKey="Lepage P">P Lepage</name>
</author>
<author><name sortKey="Harmsen, E" uniqKey="Harmsen E">E Harmsen</name>
</author>
<author><name sortKey="Hudson, Tj" uniqKey="Hudson T">TJ Hudson</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Fogarty, Mp" uniqKey="Fogarty M">MP Fogarty</name>
</author>
<author><name sortKey="Xiao, R" uniqKey="Xiao R">R Xiao</name>
</author>
<author><name sortKey="Prokunina Olsson, L" uniqKey="Prokunina Olsson L">L Prokunina-Olsson</name>
</author>
<author><name sortKey="Scott, Lj" uniqKey="Scott L">LJ Scott</name>
</author>
<author><name sortKey="Mohlke, Kl" uniqKey="Mohlke K">KL Mohlke</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Pinsonneault, Jk" uniqKey="Pinsonneault J">JK Pinsonneault</name>
</author>
<author><name sortKey="Papp, Ac" uniqKey="Papp A">AC Papp</name>
</author>
<author><name sortKey="Sadee, W" uniqKey="Sadee W">W Sadee</name>
</author>
</analytic>
</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<affiliations><list><country><li>Australie</li>
<li>France</li>
<li>États-Unis</li>
</country>
<region><li>Auvergne-Rhône-Alpes</li>
<li>Rhône-Alpes</li>
<li>Utah</li>
<li>Victoria (État)</li>
</region>
<settlement><li>Lyon</li>
<li>Melbourne</li>
</settlement>
<orgName><li>Université de Melbourne</li>
</orgName>
</list>
<tree><country name="France"><region name="Auvergne-Rhône-Alpes"><name sortKey="Nguyen Dumont, Tu" sort="Nguyen Dumont, Tu" uniqKey="Nguyen Dumont T" first="Tú" last="Nguyen-Dumont">Tú Nguyen-Dumont</name>
</region>
<name sortKey="Forey, Nathalie" sort="Forey, Nathalie" uniqKey="Forey N" first="Nathalie" last="Forey">Nathalie Forey</name>
<name sortKey="Jordheim, Lars P" sort="Jordheim, Lars P" uniqKey="Jordheim L" first="Lars P" last="Jordheim">Lars P. Jordheim</name>
<name sortKey="Le Calvez Kelm, Florence" sort="Le Calvez Kelm, Florence" uniqKey="Le Calvez Kelm F" first="Florence" last="Le Calvez-Kelm">Florence Le Calvez-Kelm</name>
<name sortKey="Lesueur, Fabienne" sort="Lesueur, Fabienne" uniqKey="Lesueur F" first="Fabienne" last="Lesueur">Fabienne Lesueur</name>
<name sortKey="Mckay Chopin, Sandrine" sort="Mckay Chopin, Sandrine" uniqKey="Mckay Chopin S" first="Sandrine" last="Mckay-Chopin">Sandrine Mckay-Chopin</name>
<name sortKey="Michelon, Jocelyne" sort="Michelon, Jocelyne" uniqKey="Michelon J" first="Jocelyne" last="Michelon">Jocelyne Michelon</name>
<name sortKey="Sinilnikova, Olga" sort="Sinilnikova, Olga" uniqKey="Sinilnikova O" first="Olga" last="Sinilnikova">Olga Sinilnikova</name>
</country>
<country name="Australie"><region name="Victoria (État)"><name sortKey="Southey, Melissa C" sort="Southey, Melissa C" uniqKey="Southey M" first="Melissa C" last="Southey">Melissa C. Southey</name>
</region>
</country>
<country name="États-Unis"><region name="Utah"><name sortKey="Tavtigian, Sean V" sort="Tavtigian, Sean V" uniqKey="Tavtigian S" first="Sean V" last="Tavtigian">Sean V. Tavtigian</name>
</region>
</country>
</tree>
</affiliations>
</record>
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