相关实验视频

Updated: Jun 19, 2026

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Published on: August 25, 2018

在难以转移的红状腺细胞系中,有效的CRISPR-Cas9介导光标签 knockin的协议

Virginie Deleuze¹, Eric Soler¹, Charlotte Andrieu-Soler¹

¹IGMM University Montpellier, CNRS, Montpellier, France; Laboratory of Excellence GR-Ex, Université' de Paris, Paris, France.

|April 19, 2024

概括

这项研究详细介绍了一种CRISPR-Cas9方法,用于将光标签插入哺乳动物基因. 它提供了一个高效的基因编辑协议,在具有挑战性的红状腺细胞系中进行编辑,帮助基因功能研究.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
细胞生物学细胞生物学

背景情况:

克里斯普尔-Cas9技术能够精确地进行基因编辑.
同源重组 (HR) 对于向基因修改至关重要.
研究基因功能需要可靠的基因标记方法.

研究的目的:

介绍一个有效的HR介导插入光标记物到哺乳动物基因的协议.
为了优化难以转移的红状腺细胞系 (MEL和MEDEP) 的基因编辑.
通过精确的基因组标记,促进对基因功能和蛋白质产品的研究.

主要方法:

针对性基因插入的CRISPR-Cas9介导的同源重组 (HR).
开发一个用于等离子体构造和电穿孔的协议.
对基因组编辑进行放大和验证的方法,包括克隆隔离和验证.

主要成果:

在MEL和MEDEP细胞系中展示HR介导的光标记物的高效插入.
在具有挑战性的红色素原生细胞中成功编辑基因组的详细步骤.
确立了识别和验证成功克隆诺基因的程序.

结论:

提出的协议使光标签能够有效地插入难以转移的红状腺细胞系.
这种方法对于研究哺乳动物系统中的基因功能和蛋白质产品非常有价值.
该协议为特定细胞环境中的基因组编辑提供了强大的方法.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?细胞隔离细胞的隔离.流动细胞计量 (Flow Cytometry) 是一种流动细胞计.基因组学就是基因组学.分子生物学分子生物学

更多相关视频

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

Published on: November 13, 2021

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

Published on: January 26, 2024

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

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...