Supplementary Material for: Cytogenomic Characterization of a Novel de novo Balanced Reciprocal Translocation t(1;12) by Genome Sequencing Leading to Fusion Gene Formation of <b><i>EYA3/EFCAB4b</i></b>

<b><i>Introduction:</i></b> The accurate detection of breakpoint regions of disease-associated chromosomal rearrangements helps understand the molecular mechanisms and identify the risks involved with disrupted genes. <b><i>Methods:</i></b> In this study, a girl with growth retardation is characterized using positional cloning and genome sequencing. The techniques include fluorescence in situ hybridization (FISH) with paint (WCP) and bacterial-artificial chromosomes (BAC) probes, PCR, real-time PCR, and short and long-read sequencing. <b><i>Results:</i></b> The translocation was identified by GTG banding and confirmed by WCP FISH. Microarray ruled out the involvement of other copy number variations except for 6 homozygous regions which are not disease-causing variants. Fine mapping with FISH showed split signals with BAC clone RP11-312A3. Genome sequencing of short-read with an average 30× depth and long-read sequencing technology with a 3.8× coverage identified both breakpoints, confirmed by Sanger sequencing, that showed microhomology. The breakpoint at 1p and 12p regions disrupted <i>EYA3</i> and <i>EFCAB4B</i> genes. Expression analysis of <i>EYA3</i> showed a 7-fold increase, suggesting the formation of a fusion gene with <i>EFCAB4B</i>. <i>EYA3</i> is involved in skeleton development, and <i>EFCAB4B</i> plays a role in calcium metabolism, which may be relevant for the patient’s phenotype. <b><i>Conclusion:</i></b> The systematic application of genome techniques to translocations and their advantages is discussed.

**<i>引言：</i>** 准确检测疾病相关染色体重排的断裂点区域，有助于解析其分子机制，并明确受累基因相关的风险。**<i>方法：</i>** 本研究通过定位克隆与基因组测序技术，对1例生长迟缓女童进行表型特征分析。所用技术包括：涂染型荧光原位杂交（Fluorescence in situ hybridization, FISH，即全染色体涂染探针WCP）、细菌人工染色体（Bacterial-artificial Chromosomes, BAC）探针、聚合酶链式反应（Polymerase Chain Reaction, PCR）、实时荧光定量PCR（real-time PCR），以及短读长测序与长读长测序技术。**<i>结果：</i>** 本研究通过GTG显带技术鉴定出该染色体易位，并经WCP-FISH验证。基因芯片检测排除了其他拷贝数变异的参与，仅检出6个非致病性纯合区域。通过荧光原位杂交进行精细定位，观察到BAC克隆RP11-312A3呈现分裂信号。采用平均测序深度为30×的短读长测序技术，以及覆盖度为3.8×的长读长测序技术，成功定位两条染色体的断裂点，经桑格测序（Sanger sequencing）验证，该断裂位点存在微同源序列。位于1p与12p区域的断裂分别破坏了*EYA3*与*EFCAB4B*基因。对*EYA3*的表达分析显示其表达量上调7倍，提示其与*EFCAB4B*形成融合基因。*EYA3*参与骨骼发育过程，而*EFCAB4B*在钙代谢中发挥作用，这可能与患者的临床表型相关。**<i>结论：</i>** 本研究探讨了基因组技术在染色体易位检测中的系统化应用及其技术优势。

搜集汇总

背景与挑战

背景概述

该数据集是《细胞基因组学特征研究：通过基因组测序表征新型从头平衡相互易位t(1;12)导致EYA3/EFCAB4b融合基因形成》的补充材料，包含图表和表格文件。研究使用基因组测序（包括短读长和长读长技术）分析了一个生长迟缓病例，识别出染色体1p和12p区域的易位断点，这些断点破坏了EYA3和EFCAB4B基因，并观察到EYA3表达增加，表明融合基因形成可能与患者表型相关。数据集突出了基因组技术在染色体易位研究中的应用价值。