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在海洋远细胞系中对CRISPR/Cas9传递方法的比较分析

Álvaro J Arana¹, Sara Veiga-Rua¹, Diego Cora²

¹Departamento de Zooloxía, Xenética e Antropoloxía Física, Facultade de Veterinaria, Universidade de Santiago de Compostela, 27002 Lugo, Spain.

International journal of molecular sciences

|November 13, 2025

概括

由于交付问题,在海洋鱼细胞系中进行CRISPR/Cas9基因编辑具有挑战性. 这项研究发现,电穿孔在SaB-1细胞中效果最好,但输送方法取决于细胞系和细胞内因素.

科学领域:

海洋生物学海洋生物学
基因组学就是基因组学.
生物技术是生物技术.

背景情况:

CRISPR/Cas9基因编辑为功能基因组学提供了强大的工具.
有效地将CRISPR/Cas9组件输入海洋鱼细胞系是一个重大障碍.
了解交付机制对于推进海洋远洋研究和水产养殖至关重要.

研究的目的:

在海洋鱼细胞系中比较三种CRISPR/Cas9传递方法 (电穿孔,脂质纳米粒子,磁传染) 的效率.
评估传递方法对转染效率,编辑结果和基因组稳定性的影响.
确定影响DLB-1和SAB-1细胞系成功基因编辑的因素.

主要方法:

在DLB-1和SaB-1细胞系中进行了CRISPR/Cas9介导的ifi27l2a基因编辑.
测试了三个输送策略:电穿孔,脂质纳米粒子 (LNPs) 和使用SPIONs的磁传染.
评估了传染效率,编辑速率,Cas9细胞内定位和基因组稳定性.

主要成果:

在 SaB-1 细胞中,电穿孔实现了高编辑效率 (高达 95%),但在 DLB-1 细胞中,基因组重组时,效率较低 (30%).
脂质纳米颗粒在DLB-1细胞中显示中度编辑 (~25%),但在SaB-1细胞中编辑最小.
基于SPION的磁传感导致了有效的粒子吸收,但没有可检测的编辑,表明进入后的障碍.
Cas9细胞内流通和聚合影响了编辑成功.

结论:

在海洋远程生物中,CRISPR/Cas9传递效率高度依赖细胞系.
细胞内贩运,Cas9定位和基因组完整性是成功基因编辑的关键因素.
这些发现为优化海洋鱼类基因编辑策略的研究和水产养殖提供了一个框架.

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.水产养殖的水产养殖编辑效率编辑效率编辑效率基因编辑基因编辑海鱼是什么意思海鱼是什么意思海 (Sea Bream) 是一种海.

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