Generation of isogenic models of Angelman syndrome and Prader-Willi syndrome in CRISPR/Cas9-engineered human embryonic stem cells.
Generation of isogenic models of Angelman syndrome and Prader-Willi syndrome in CRISPR/Cas9-engineered human embryonic stem cells.
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在 CRISPR/Cas9 工程人类胚胎干细胞中生成 Angelman 综合征和 Prader-Willi 综合征的等基因模型。
DOI：
10.1101/2023.08.30.555563
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Chamberlain,StormyJ
中科院分区：
文献类型：
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作者：
Gilmore,RachelB;Gorka,Dea;Stoddard,ChristopherE;Cotney,JustinL;Chamberlain,StormyJ
Angelman综合征(AS)和Prader-Willi综合征(PWS)是两种不同的神经发育障碍，由15q11-13基因座印记基因表达缺失引起，最常见的原因是母亲或父亲的等位基因大规模缺失。每一例的发病率约为1/15,000至1/30,000活产，并具有一系列使人衰弱的表型。患者诱导的多能干细胞(IPSCs)已经成为了解该基因座人类相关基因调控的宝贵工具，并为AS治疗方法的发展做出了贡献。然而，对于这些缺失如何导致AS和PWS的失调和表型，我们的理解仍然存在差距。由于供体差异、重新编程方法和遗传背景导致的细胞系之间的可变性，使得在不大幅增加分析中使用的细胞系数量的情况下填补这些知识空白具有挑战性。只有引起疾病的基因突变不同的同基因细胞系可以减轻这一负担，而不需要如此大量的细胞系。在这里，我们描述了模拟AS和PWS最常见的遗传亚型的等基因人类胚胎干细胞(HESC)系的发展。这些品系可以方便地询问染色体15q11-q13基因座上的等位基因特异性基因调控。此外，这些品系是确定和测试AS和PWS患者靶向治疗方法的重要资源。
Angelman syndrome (AS) and Prader-Willi syndrome (PWS), two distinct neurodevelopmental disorders, result from loss of expression from imprinted genes in the chromosome 15q11-13 locus most commonly caused by a megabase-scale deletion on either the maternal or paternal allele, respectively. Each occurs at an approximate incidence of 1/15,000 to 1/30,000 live births and has a range of debilitating phenotypes. Patient-derived induced pluripotent stem cells (iPSCs) have been valuable tools to understand human-relevant gene regulation at this locus and have contributed to the development of therapeutic approaches for AS. Nonetheless, gaps remain in our understanding of how these deletions contribute to dysregulation and phenotypes of AS and PWS. Variability across cell lines due to donor differences, reprogramming methods, and genetic background make it challenging to fill these gaps in knowledge without substantially increasing the number of cell lines used in the analyses. Isogenic cell lines that differ only by the genetic mutation causing the disease can ease this burden without requiring such a large number of cell lines. Here, we describe the development of isogenic human embryonic stem cell (hESC) lines modeling the most common genetic subtypes of AS and PWS. These lines allow for a facile interrogation of allele-specific gene regulation at the chromosome 15q11-q13 locus. Additionally, these lines are an important resource to identify and test targeted therapeutic approaches for patients with AS and PWS.