Trichostatin A for Efficient CRISPR-Cas9 Gene Editing of Human Pluripotent Stem Cells.
Trichostatin A for Efficient CRISPR-Cas9 Gene Editing of Human Pluripotent Stem Cells.
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DOI：
10.1089/crispr.2023.0033
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发表时间：
2023-09
期刊：
影响因子：
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通讯作者：
Kaivalya Molugu;Namita Khajanchi;C. Lazzarotto;S. Tsai;Krishanu Saha
中科院分区：
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作者：
Kaivalya Molugu;Namita Khajanchi;C. Lazzarotto;S. Tsai;Krishanu Saha
基因组编辑的人诱导多能干细胞(IPSCs)在疾病建模、药物开发和再生医学中有着广泛的应用。尽管发展了簇状规则间隔短回文重复序列(CRISPR)-Cas9系统，但基因编辑过程效率低下，可能需要几周到几个月的时间才能产生编辑后的IPSC克隆。我们开发了一种策略，通过应用小分子曲古抑素A(TSA)来提高IPSC基因编辑过程的效率，TSA是一种I类和II类组蛋白去乙酰化酶抑制剂。我们观察到，TSA减少了整体染色质的凝聚，并进一步导致IPSCs的基因编辑效率提高了两到四倍，同时确保不会增加非靶标效应。编辑后的IPSCs可以在保持基因组完整性和多能性的同时进行克隆扩增。这些发现可能使治疗相关基因编辑的ipscs的快速生成成为可能。
Genome-edited human-induced pluripotent stem cells (iPSCs) have broad applications in disease modeling, drug discovery, and regenerative medicine. Despite the development of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system, the gene editing process is inefficient and can take several weeks to months to generate edited iPSC clones. We developed a strategy to improve the efficiency of the iPSC gene editing process via application of a small-molecule, trichostatin A (TSA), a Class I and II histone deacetylase inhibitor. We observed that TSA decreased global chromatin condensation and further resulted in increased gene-editing efficiency of iPSCs by twofold to fourfold while concurrently ensuring no increased off-target effects. The edited iPSCs could be clonally expanded while maintaining genomic integrity and pluripotency. The rapid generation of therapeutically relevant gene-edited iPSCs could be enabled by these findings.