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通过使用Cas9 (VULSCi002-A-2) 来生成一个基因编码的电压指示器 MARINA reporter人类iPS细胞系

Aistė Petruškevičiūtė¹, Ugnė Šimuliūnaitė², Catalina M Polanco¹

¹VU LSC - EMBL Partnership Institute for Gene Editing Technologies, Saulėtekio avenue 7, Vilnius, Lithuania; Laboratory of Nuclease Enabled Cell Therapies, Lithuania.

Stem cell research

|December 25, 2024

概括

像MARINA这样的基因编码电压指示器 (GEVI) 能够对细胞膜潜力的高通量选. 研究人员开发了人类诱导的多能干细胞系,用于高级细胞研究.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

遗传编码电压指示器 (GEVI) 对于监测细胞膜潜力至关重要.
玛丽娜,从ArcLight衍生出的GEVI,与其前身不同,表现出增加的光与脱极化.
人类诱导的多能干细胞 (hiPSCs) 为细胞研究提供了有价值的模型.

研究的目的:

创建和描述人类iPSC线条,其中包括MARINA GEVI记者.
建立一个用于膜潜在表型的高通量选的工具.
在人类细胞环境中研究 MARINA 的实用性.

主要方法:

使用CRISPR-Cas9基因编辑 (SpCas9) 进行精确的MARINA记者敲入.
产生异质合体的 knock-in 记者人类 iPSC 线.
在iPSC线路中描述了MARINA记者的表情和功能.

主要成果:

成功生成了人类iPSC线条,将MARINA GEVI集成到基因组中.
在iPSC克隆中展示了MARINA记者的成功表达.
使用 MARINA 建立了研究人类细胞膜潜力动态的基础.

结论:

开发的MARINA表达的hiPSC线是神经科学和药物发现的一个有前途的工具.
这个记者系统促进了对细胞电生理学的先进研究.
玛丽娜的独特特性为复杂的生物系统中的膜潜力监测提供了优势.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?这就是Cas9的情况.有基因编码的记者玛丽娜·马里纳这些细胞是iPS细胞.

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