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在分化后的人类iPSC心肌细胞中,CRISPRi基因调制和全光电生理学

Julie L Han¹, Yuli W Heinson¹, Christianne J Chua¹

¹Department of Biomedical Engineering, The George Washington University, Washington, DC, 20052, USA.

Communications biology

|December 7, 2023

概括

克里斯普尔干扰 (CRISPRi) 有效地修改了从干细胞衍生的人类心脏细胞中的基因功能. 这一进展有助于理解基因与疾病的联系,并开发新的心脏病治疗方法.

科学领域:

心脏病学心脏病学
基因组学就是基因组学.
干细胞生物学干细胞生物学

背景情况:

在人类诱导的多能干细胞衍生心肌细胞 (iPSC-CMs) 中精确的基因调制对于理解基因-表型关系至关重要.
在功能性基因组学中,CRISPR衍生技术提供可逆的基因抑制或激活 (CRISPRi/a).

研究的目的:

在分化后的iPSC-CM中描述CRISPR干扰 (CRISPRi) 性能.
评估CRISPRi对心脏离子通道基因 (KCNH2,KCNJ2,GJA1) 的影响.
用全光电生理学量化对心脏再极化,静止膜电位和导电的影响.

主要方法:

在差异化的iPSC-CM中利用了CRISPRi技术.
有针对性的关键心脏离子通道基因:KCNH2,KCNJ2和GJA1.
采用全光电生理学用于多参数表型化.

主要成果:

优化CRISPRi传递 (例如Zim3效应器,病毒载体) 在非分裂细胞中提高了性能.
实现了与CRISPRi iPSC系列相提并论的性能.
在差异化的iPSC-CM中显示出轻度但特定的基因敲除表型.

结论:

克里斯皮尔是有效调节基因功能在差异化的iPSC-CMs.
这种方法促进了临床前心脏毒性测试.
潜在的未来 in vivo 治疗应用.

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