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使用CRISPR/Cas9系统在哺乳动物细胞系中生成单细胞衍生淘汰克隆的指南

Taeyeon Hong¹, Seung-Min Bae², Gwonhwa Song³

¹Department of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 16419, Republic of Korea.

Molecules and cells

|June 27, 2024

概括

这项研究指导研究人员使用聚类定期间隔的短巴林德罗姆重复相关蛋白9 (CRISPR-Cas9) 进行精确的基因组编辑. 它详细介绍了产生淘汰细胞系的方法,并为人类和动物细胞研究中的应用提供了洞察力.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

基因组编辑可以在各种生物体中进行有针对性的修改.
聚类定期间隔的短巴林德罗姆重复相关蛋白9 (CRISPR-Cas9) 系统是精确基因编辑的强大工具.
在基因疾病纠正和癌症治疗方面,CRISPR-Cas9具有显著的临床潜力.

研究的目的:

为利用CRISPR-Cas9技术在研究环境中提供全面指南.
详细介绍生成转基因细胞系和动物模型的方法.
为研究哺乳动物细胞系的研究人员提供实用信息.

主要方法:

单导向RNA (sgRNA) 设计和克隆成等离子体载体.
转染后单细胞分离. 转染后单细胞分离.
使用下一代测序 (NGS) 识别淘汰克隆.
通过NGS.对哺乳动物细胞系中基因插入的分析.

主要成果:

在哺乳动物细胞系中成功生成淘汰克隆.
通过下一代测序验证基因编辑效率.
在细胞系修改中展示了CRISPR-Cas9的实际应用.

结论:

这项研究为CRISPR-Cas9介导的基因组编辑提供了一个有价值的协议.
这些发现支持使用CRISPR-Cas9来在研究中制造精确的基因修饰.
这项工作为人类和动物细胞系研究的研究人员提供了实用数据.

关键词:

聚类有规律间隔的短巴林德罗姆重复-关联蛋白 9 9 文化系统文化系统基因组编辑基因组编辑单导向RNA是一种单导向RNA.

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