Parental-origin-determination fluorescence in situ hybridization distinguishes homologous human chromosomes on a single-cell level.
Parental-origin-determination fluorescence in situ hybridization distinguishes homologous human chromosomes on a single-cell level.
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亲本来源测定荧光原位杂交可在单细胞水平上区分同源人类染色体。
DOI：
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发表时间：
2008
影响因子：
5.4
通讯作者：
T. Liehr
中科院分区：
文献类型：
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作者：
A. Weise;M. Gross;K. Mrasek;H. Mkrtchyan;Bernhard Horsthemke;C. Jonsrud;F. Eggeling;Sophie Hinreiner;Vera Witthuhn;U. Claussen;T. Liehr
同源染色体的分化以及它们的亲本来源目前可以通过使用微卫星或SNP分析的分子遗传学方法进行和专门确定。只有在例外情况下，才可能在单细胞水平上进行区分，例如由于1号、9号、16号和Y号染色体的异染色质区域或近端着丝粒染色体的p臂内的变异。在缺乏这种多态性的情况下，同源染色体的个体区分目前是不可能的。因此，各种科学和诊断相关的问题无法得到回答。基于最近检测到的大规模拷贝数变异（LCV）或拷贝数多态性（CNP），在本研究中，提出了一种用于同源染色体个体间分化的分子细胞遗传学技术，称为亲本来源确定荧光原位杂交（pod-FISH）。所有人类染色体均覆盖有225个LCV-和/或CNP-特异性BAC探针，并创建、评估和优化一至五色染色体特异性pod-FISH集。我们证明，pod-FISH适用于单细胞分析单亲二体性（UDP）在临床病例中，如普拉德-威利综合征引起的母亲UPD。一个罕见的临床病例与马赛克形式的全基因组isodisomy被用来确定检测限的pod-FISH。此外，我们首次分析了常规微卫星分析的信息量，并将结果与pod-FISH进行了比较。有了这种在单细胞水平上研究个体人类染色体的亲本起源的新可能性，为诊断和基础研究打开了新的大门。
The differentiation of homologous chromosomes as well as their parental origin can presently be conducted and determined exclusively by molecular genetic methods using microsatellite or SNP analysis. Only in exceptional cases is a distinction on a single-cell level possible, e.g. due to variations within the heterochromatic regions of chromosomes 1, 9, 16 and Y or the p-arms of the acrocentric chromosomes. In the absence of such polymorphisms, an individual distinction of the homologous chromosomes is not currently possible. Consequently, various questions of scientific and diagnostic relevance are unable to be answered. Based on the recently detected large-scale copy-number variations (LCV) or copy-number polymorphisms (CNP) spanning up to several megabase pairs of DNA, in this study, a molecular cytogenetic technique for the inter-individual differentiation of homologous chromosomes called parental-origin-determination fluorescence in situ hybridization (pod-FISH) is presented. All human chromosomes were covered with 225 LCV- and/or CNP-specific BAC probes, and one- to five-color chromosome-specific pod-FISH sets were created, evaluated and optimized. We demonstrated that pod-FISH is suitable for single-cell analysis of uniparental disomy (UDP) in clinical cases such as Prader-Willi syndrome caused by maternal UPD. A rare clinical case with a mosaic form of a genome-wide isodisomy was used to determine the detection limits of pod-FISH. Additionally we analyzed the informativeness of conventional microsatellite analysis for the first time and compared the results to pod-FISH. With this new possibility to study the parental origin of individual human chromosomes on a single-cell level, new doors for diagnostic and basic research are opened.
影响因子：
9.8
作者：
Locke, Devin P.;Sharp, Andrew J.;Eichler, Evan E.
通讯作者：
Eichler, Evan E.