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基于病毒报告基因组合的转录因子活性的PCR分析

PCR-based profiling of transcription factor activity by a virus-based reporter battery.

作者信息

Abe Hitomi, Abe Kentaro

机构信息

Lab of Brain Development, Graduate School of Life Sciences, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai, Miyagi 980-8577, Japan.

Division for the Establishment of Frontier Sciences, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai, Miyagi 980-8577, Japan.

出版信息

iScience. 2022 Feb 14;25(3):103927. doi: 10.1016/j.isci.2022.103927. eCollection 2022 Mar 18.

Abstract

Understanding the molecular mechanisms of gene regulation is pivotal for understanding how cells establish and modify their identities and functions. Multiple transcription factors (TFs) coordinate to alter gene expression in cells; however, a method to quantitatively analyze the activity of each TF is lacking, particularly . Here, we introduce a viral-vector-based TF reporter battery that can be used to simultaneously analyze the activity of multiple TFs, visualized as the TF activity profile (TFAP) obtained by qPCR. We show that the cells possess distinct TFAPs that dynamically change according to experimental manipulation or physiological activity. We report a practical method to obtain the TFAP of a defined cell population and their experience-dependent changes in the mouse brain The TFAP obtained by our method will help bridge the information gap between the genome and transcriptome and aid the multi-omics view of understanding the gene regulation system.

摘要

理解基因调控的分子机制对于理解细胞如何建立和改变其特性及功能至关重要。多种转录因子(TFs)协同作用以改变细胞中的基因表达;然而,目前缺乏一种定量分析每个TF活性的方法,尤其是……在这里,我们引入了一种基于病毒载体的TF报告基因组合,可用于同时分析多个TF的活性,通过qPCR获得的TF活性谱(TFAP)进行可视化。我们表明,细胞具有独特的TFAP,其会根据实验操作或生理活动而动态变化。我们报告了一种实用的方法,可在小鼠大脑中获得特定细胞群体的TFAP及其依赖经验的变化。通过我们的方法获得的TFAP将有助于弥合基因组和转录组之间的信息差距,并有助于从多组学角度理解基因调控系统。

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