相关实验视频

Updated: Jun 13, 2025

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

Published on: March 27, 2020

使用条形码表达式记者的CRISPRi在人类细胞中剖析了调节网络

Jinyoung Kim¹, Ryan Y Muller¹, Eliana R Bondra¹

¹Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.

bioRxiv : the preprint server for biology

|September 16, 2024

概括

我们开发了CiBER-seq,一种使用CRISPR干扰 (CRISPRi) 和条码测序来测量基因表达变化的方法. 这种技术准确地绘制了人类细胞中的遗传网络和转录反应,具有高时间分辨率.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
细胞生物学细胞生物学

背景情况:

全基因组的CRISPR屏幕对于理解基因功能至关重要.
测量分子表现型,如基因表达,是有效选的关键.
与报告员系统相结合的CRISPR干扰 (CRISPRi) 为定量分析提供了一条道路.

研究的目的:

引入和验证CiBER-seq,一种用于量化转录反应的新方法.
展示CiBER-seq在剖析复杂遗传网络中的应用.
优化CiBER-seq用于具有大规模扰动库的哺乳动物细胞.

主要方法:

将CRISPR干扰 (CRISPRi) 与条码表达式记者测序 (CiBER-seq) 结合起来.
优化将复杂的条形码sgRNA库集成到定义的基因组环境中.
使用大量RNA测序来捕获快速的转录反应.

主要成果:

CiBER-seq准确地测量了跨基因组规模扰乱库的转录反应.
对核因子kappa B (NF-κB) 记者进行分析,揭示了TNF-alpha信号级联.
确定了参与炎症基因激活的细胞类型特定因素.

结论:

CiBER-seq是一个准确而强大的工具,用于剖析哺乳动物细胞中的遗传网络.
该方法使得研究快速转录事件的时间分辨率更高.
CiBER-seq促进了分子表型的定量测量,以应对遗传干扰.

更多相关视频

Author Spotlight: An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

Author Spotlight: An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

Published on: April 21, 2023

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Published on: March 7, 2018

相关概念视频

Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.