相关实验视频

Updated: May 16, 2026

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

使用CRISPR/Cas9生成难以转移的THP1细胞系中单基因淘汰的协议

Kaveri Srivastava^1,2, Bhaswati Pandit^1,2

¹BRIC-National Institute of Biomedical Genomics (BRIC-NIBMG) Kalyani, West Bengal, India.

|July 14, 2025

概括

该协议详细介绍了一种CRISPR-Cas9方法,用于在难以转移的免疫细胞 (如THP1.1) 中创建单基因淘汰. 它使用lentiviral交付进行高效的基因编辑,使功能研究.

科学领域:

免疫学免疫学免疫学
分子生物学分子生物学
遗传学是一个遗传学.

背景情况:

免疫细胞系,如THP1,对于研究细胞通路至关重要,但往往很难进行基因修饰.
有效的基因淘汰方法对于理解这些复杂系统中的基因功能至关重要.

研究的目的:

建立一个可靠的协议,在难以转移的悬浮免疫细胞中产生单基因淘汰.
通过透视病毒传递的CRISPR-Cas9系统在THP1细胞中精确地破坏基因的有效性.

主要方法:

设计和克隆单导向RNAs (sgRNAs) 进入CRISPR矢量.
病毒生产和转导THP1细胞用于基因传递.
基因淘汰的选择和逐步验证使用殖民地PCR,测序和西式涂抹.

主要成果:

在THP1细胞中成功生成单基因淘汰 (GSDMD).
在悬浮免疫细胞中的基因编辑中,lentiviral的输送显示出高效率.
综合验证证实了淘汰赛协议的准确性和可靠性.

结论:

这种优化的协议提供了一种可扩展和有效的方法,用于在具有挑战性的免疫细胞系中进行CRISPR-Cas9介导的基因淘汰.
该协议通过在免疫细胞中实现精确的基因破坏和随后的分析来促进功能基因组研究.

关键词:

这就是CRISPR/Cas9的作用.GSDMDMD的总体发展目标是什么基因淘汰赛基因淘汰赛林氏病毒 (Lentivirus) 是一种隐形病毒.在 THP1 细胞中.

更多相关视频

Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System

Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System

Published on: April 11, 2025

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Published on: February 28, 2025

相关概念视频

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...