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Updated: Sep 3, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Published on: September 4, 2019

在整个生物体中大规模的多重化马赛克乱

Bo Liu¹, Zhengyu Jing¹, Xiaoming Zhang¹

¹Gene Editing Center, School of Life Sciences and Technology, ShanghaiTech University, Shanghai 201210, China.

|July 23, 2022

概括

我们开发了一种可诱导的动态动态模型 (iMAP), 这种系统可以同时针对整个身体的基因, 显示特定的基因功能.

科学领域:

遗传学
分子生物学
系统生物学

背景情况:

了解组织特异性的基因功能对于解读复杂的生物过程至关重要.
现有的大规模遗传干扰方法往往缺乏在多种组织中进行现场分析的分辨率.
开发先进的转基因平台对于全面的基因组解码至关重要.

研究的目的:

引入可诱导的动植物扰动 (iMAP) 系统,用于小鼠的高通量现场遗传查.
证明iMAP能够同时针对整个生物体中的许多基因.
创建一个初步的Perturb-Atlas以识别上下文依赖的基因功能.

主要方法:

使用Cre-loxP和CRISPR-Cas9技术开发一个转基因小鼠模型 (iMAP).
一个转基因的生殖线传输编码单独流动的gRNA单元的串联阵列.
在各种组织中诱导单个gRNA的马赛克表达.
使用gRNA表示用于表型特征和基因扰乱映射.

主要成果:

已经成功生成了能够同时向至少100个基因的马赛克小鼠.
在39个不同的小鼠组织中绘制了90个基因的扰动, 创建了一个微型的 Perturb-Atlas.
通过组织特异性扰动分析对上下文依赖的基因功能的新见解.
证明iMAP对单基因扰动线的高通量导出具有实用性.

结论:

iMAP平台为现场,全身基因扰动和功能基因组学提供了强大的工具.
iMAP促进了特定环境的基因角色的发现,并加速了基因组解码工作.
这项技术有助于我们更好地理解复杂生物系统中的基因功能.

关键词:

汽车T 波动后的时间细胞地图免疫疗法在体内进行CRISPR检查小鼠细胞 Perturb-Atlas

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