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使用CRISPR/Cas9技术对纤维瘤细胞中AKT1淘汰的功能分析

So-Hee Kim¹, Moon-Moo Kim²

¹Department of Cosmetics Engineering, and Graduate School of Applied Chemistry·Food Engineering, Dong-Eui University, Busan, Republic of Korea.

Journal of human genetics

|November 26, 2025

概括

这项研究使用CRISPR/Cas9生成了AKT1淘汰细胞,以了解AKT1基因在细胞信号传输中的功能. AKT1中断显著改变了下游效应器表达,揭示了它的调节作用.

科学领域:

分子生物学分子生物学
细胞信号传输细胞信号传输
基因编辑基因编辑

背景情况:

AKT1 (蛋白激酶Bα) 是一个关键的氨酸/氨酸激酶,调节细胞过程.
了解AKT1在信号通路中的作用对于细胞功能研究至关重要.

研究的目的:

阐明AKT1基因在细胞信号通路中的作用.
使用CRISPR/Cas9技术生成AKT1淘汰 (KO) 的HT-1080细胞.

主要方法:

克里斯普尔/卡斯9系统用于AKT1基因编辑.
桑格DNA测序和InDel分析用于验证.
定量实时PCR和西布洛特用于表达式分析.

主要成果:

通过验证基因编辑成功生成了AKT1 KO HT-1080细胞.
在KO细胞中显著减少AKT1mRNA和蛋白质表达.
在下游效应因子:mTOR,BCL-2和FOXO1.1中显示出显著的变化.

结论:

破坏AKT1基因会影响关键的细胞信号通路.
提供了关于AKT1基因调控功能的基本见解.
已建立的AKT1KO细胞作为进一步研究的宝贵工具.

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