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Updated: Jul 13, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

基因造型 - - CRISPR局部编辑的多重测序 (GMUSCLE):一种用于分析CRISPR编辑细胞的实验和计算方法

Peng Zhang¹, Laurent Abel^1,2,3, Jean-Laurent Casanova^1,2,3,4,5

¹St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, New York, USA; Necker Hospital for Sick Children, Paris, France.

The CRISPR journal

|October 12, 2023

概括

基因型定型聚类定期间隔短帕林德罗姆重复-CRISPR相关蛋白9 (CRISPR-Cas9) 编辑的细胞通过新协议和GMUSCLE软件简化. 这种方法可以有效和准确地识别所需的细胞基因型.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物信息学是一种生物信息学.

背景情况:

克里斯普尔-Cas9基因编辑会诱导双链断裂,导致细胞间异质和细胞间异质.
基因定型CRISPR-Cas9编辑细胞是复杂的,传统的方法是耗时的.
有效和准确的基因造型对于选择和分析编辑细胞克隆至关重要.

研究的目的:

开发一种精简的基因定型CRISPR-Cas9编辑细胞协议.
引入用于定量和定性基因型识别的GMUSCLE软件.
为了实现成本效益高的多重测序,用于CRISPR-Cas9编辑分析.

主要方法:

开发了一个精简的CRISPR-Cas9基因定型的实验和计算协议.
利用成本效益高的多重测序来进行编辑产品的深度测序.
创建了用于分析测序数据和识别基因型的GMUSCLE软件.
验证了使用CRISPR-Cas9干扰干扰素α/β受体alpha的协议和软件.

主要成果:

成功验证了用于基因定型CRISPR-Cas9编辑细胞的协议和GMUSCLE软件.
同时确定了20个通过CRISPR-Cas9.9编辑的细胞克隆的基因型.
证明了GMUSCLE能够分析来自单个克隆和批量细胞群的测序数据的能力.
实现了定量和定性基因型识别,促进了具有特定基因型的克隆的选择.

结论:

新的协议和GMUSCLE软件显著简化了CRISPR-Cas9编辑细胞的基因定型.
这种方法可以有效地选择和研究具有所需基因型的细胞克隆.
GMUSCLE是一个多功能工具,适用于多重和批量人口测序数据.
开发的方法为CRISPR-Cas9编辑分析提供了具有成本效益和准确的解决方案.

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