Recent advances in genome engineering based on the CRISPR/Cas9 technology have revolutionized our ability to manipulate genomic DNA. Its use in human pluripotent stem cells (hPSCs) has allowed a wide range of mutant cell lines to be obtained at an unprecedented rate. The combination of these two groundbreaking technologies has tremendous potential, from disease modeling to stem cell-based therapies. However, the generation, screening and molecular characterization of these cell lines remain a cumbersome and multi-step endeavor. Here, we propose a pipeline of strategies to efficiently generate, sub-clone, and characterize CRISPR/Cas9-edited hPSC lines in the function of the introduced mutation (indels, point mutations, insertion of large constructs, deletions).

中文翻译：

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使用CRISPR / Cas9技术的转基因人类多能干细胞生成和表征管道。

基于CRISPR / Cas9技术的基因组工程的最新进展彻底改变了我们操作基因组DNA的能力。它在人类多能干细胞（hPSC）中的使用已允许以前所未有的速度获得各种​​突变细胞系。从疾病建模到基于干细胞的疗法，这两种突破性技术的结合具有巨大的潜力。然而，这些细胞系的产生，筛选和分子表征仍然是繁琐且多步骤的努力。在这里，我们提出了一系列策略，可以有效地产生，亚克隆和表征CRISPR / Cas9编辑的hPSC细胞系的功能，包括引入的突变（插入片段，点突变，插入大型构建体，缺失）。