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通过CRISPR/Cas9介导的Fel d1的CH2删除触发了猫细胞中的转录基因重编程和与疾病相关的途径

Lin-Yi Qu¹, Fu-Shi Quan¹, Shu-Ming Shi¹

¹College of Animal Sciences, Jilin University, Changchun, Jilin, China.

Frontiers in cell and developmental biology

|December 8, 2025

概括

基因编辑Fel d1,主要的猫类过敏原,成功降低了过敏原水平,但导致了意外的基因表达变化. 在培养低过敏性猫之前,全面的安全评估至关重要.

科学领域:

遗传学和分子生物学
过敏学过敏学
动物生物技术动物生物技术

背景情况:

Fel d1是主要的猫类过敏原,在人类中引起过敏反应.
Fel d1 的生理作用在很大程度上是未知的.
培养低过敏性猫是过敏患者的一个重要目标.

研究的目的:

通过敲除其CH2域来研究Fel d1的功能.
评估使用基因编辑生产低过敏性猫的可行性.
评估Fel d1基因淘汰的安全性和潜在副作用.

主要方法:

使用CRISPR/Cas9基因编辑,在猫皮肤细胞中敲除Fel d1的CH2域.
绝杀效率通过测序,qRT-PCR和西部涂抹得到证实.
使用RNA-seq,GO,KEGG和GSEA分析了转录组变化,使用CCK-8,EDU和流细胞计进行了表型评估.

主要成果:

成功生成了一个框架转移突变,导致Fel d1 mRNA和蛋白质几乎完全丧失.
RNA-seq揭示了3,469个差异表达的基因,其中丰富的基因存在于多变性心肌病和类风湿性关节炎的途径中.
缺失CH2损害了细胞增殖和亡的增加,并对TGFB2,MYBPC3,MMP3和TLR4等关键基因进行上调.

结论:

虽然CH2域删除有效地消除了主要的猫类过敏原Fel d1,但它会诱导显著的转录基因重编程.
这些意想不到的变化与病理状态有关,突出显示了潜在的安全问题.
在利用基因编辑细胞用于低过敏性猫的发育之前,包括转录组学在内的全面安全分析至关重要.

关键词:

在CH2中,CH2是二氧化碳.这就是CRISPR/Cas9的作用.在FEL d1中,过度缩性心肌病变性心脏病.类风湿性关节炎类风湿性关节炎转录基因组测序的转录基因组测序

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