Incorporation of 5-ethynyl-2'-deoxyuridine (EdU) as a novel strategy for identification of the skewed X inactivation pattern in balanced and unbalanced X-rearrangements

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dc.citation.issue2
dc.citation.volume135
dc.contributor.authorSisdelli, Luiza [UNIFESP]
dc.contributor.authorVidi, Angela Cristina [UNIFESP]
dc.contributor.authorMoyses-Oliveira, Mariana [UNIFESP]
dc.contributor.authorDi Battista, Adriana [UNIFESP]
dc.contributor.authorBortolai, Adriana [UNIFESP]
dc.contributor.authorMoretti-Ferreira, Danilo
dc.contributor.authorSilva, Magnus R. Dias da [UNIFESP]
dc.contributor.authorMelaragno, Maria Isabel [UNIFESP]
dc.contributor.authorCarvalheira, Gianna [UNIFESP]
dc.coverageNew York
dc.date.accessioned2020-10-30T18:46:35Z
dc.date.available2020-10-30T18:46:35Z
dc.date.issued2016
dc.description.abstractX-chromosome inactivation occurs randomly in normal female cells. However, the inactivation can be skewed in patients with alterations in X-chromosome. In balanced X-autosome translocations, normal X is preferentially inactivated, while in unbalanced X alterations, the aberrant X is usually inactivated. Here, we present a novel strategy to verify the skewed X inactivation pattern through the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) into cells, in 11 patients: five carriers of balanced X-autosome translocations and six of unbalanced X-chromosome alterations. Since EdU is a labeled nucleoside analog of thymidine, its incorporation during DNA synthesis can reveal late replication regions and the inactive X-chromosome. All EdU findings were validated by the human androgen receptor gene (HUMARA) assay. The late replication regions were easily and quickly visualized in all cells, where inactive Xs are marked with strong green fluorescence. It was observed that the normal X-chromosome was preferentially inactivated in patients with balanced X-autosome translocations; while the aberrant X-chromosome was inactivated in most cells from patients with unbalanced alterations. By performing the fluorescence-based EdU assay, the differences between the active and inactive X-chromosomes are more easily recognizable than by classic cytogenetic methods. Furthermore, EdU incorporation allows the observation of the late replication regions in autosomal segments present in X derivatives from X-autosome translocations. Therefore, EdU assay permits an accurate and efficient cytogenetic evaluation of the X inactivation pattern with a low-cost, easy to perform and highly reproducible technique.en
dc.description.affiliationUniv Fed Sao Paulo, Dept Morphol & Genet, BR-04023900 Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Dept Biochem, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Dept Med, Sao Paulo, Brazil
dc.description.affiliationSao Paulo State Univ, Inst Biociencias Botucatu, Dept Genet, BR-18618970 Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Morphol & Genet, BR-04023900 Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Biochem, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Med, Sao Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)
dc.format.extent185-192
dc.identifierhttps://doi.org/10.1007/s00439-015-1622-x
dc.identifier.citationHuman Genetics. New York, v. 135, n. 2, p. 185-192, 2016.
dc.identifier.doi10.1007/s00439-015-1622-x
dc.identifier.issn0340-6717
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/58521
dc.identifier.wosWOS:000368187000003
dc.language.isoeng
dc.publisherSpringer
dc.relation.ispartofHuman Genetics
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titleIncorporation of 5-ethynyl-2'-deoxyuridine (EdU) as a novel strategy for identification of the skewed X inactivation pattern in balanced and unbalanced X-rearrangementsen
dc.typeinfo:eu-repo/semantics/article
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