Modeling Disease in Human ESCs Using an Efficient BAC-Based Homologous Recombination System
Modeling Disease in Human ESCs Using an Efficient BAC-Based Homologous Recombination System
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DOI：
10.1016/j.stem.2009.11.016
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发表时间：
2010-01-08
期刊：
影响因子：
23.9
通讯作者：
Xu, Yang
中科院分区：
文献类型：
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作者：
Song, Hoseok;Chung, Sun-Ku;Xu, Yang
虽然小鼠模型对研究人类疾病很有价值，但小鼠和人类之间的细胞和生理差异使得开发更相关的人类疾病模型对机制研究和药物开发越来越重要。人类胚胎干细胞(HESCs)可以进行无限的自我更新，并保持分化为所有类型细胞的潜力，提供了一种可能的解决方案。为了提高人类胚胎干细胞的遗传操作效率，我们开发了基于细菌人工染色体(BAC)的方法，实现了高效的同源重组。通过用BAC靶向载体顺序干扰ATM或P53的两个等位基因，我们建立了ATM(-/-)和P53(-/-)hESCs作为人类两种主要遗传不稳定综合征的模型，并利用所产生的细胞来揭示P53在维持hESCs基因组稳定中的重要性。我们的发现表明，将转基因hESCs作为相关的人类疾病模型是可行的。
Although mouse models have been valuable for studying human disease, the cellular and physiological differences between mouse and human have made it increasingly important to develop more relevant human disease models for mechanistic studies and drug discovery. Human embryonic stem cells (hESCs), which can undergo unlimited self-renewal and retain the potential to differentiate into all cell types, present a possible solution. To improve the efficiency of genetic manipulation of hESCs, we have developed bacterial artificial chromosome (BAC) based approach that enables high efficiency homologous recombination. By sequentially disrupting both alleles of ATM or p53 with BAC targeting vectors, we have established ATM(-/-) and p53(-/-) hESCs as models for two major human genetic instability syndromes and used the generated cells to reveal the importance of p53 in maintaining genome stability of hESCs. Our findings suggest that it will be feasible to develop genetically modified hESCs as relevant human disease models.