High-throughput functional characterization of combinations of transcriptional activators and repressors.

IF 9 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Cell Systems Pub Date : 2023-09-20 Epub Date: 2023-08-04 DOI:10.1016/j.cels.2023.07.001
Adi X Mukund, Josh Tycko, Sage J Allen, Stephanie A Robinson, Cecelia Andrews, Joydeb Sinha, Connor H Ludwig, Kaitlyn Spees, Michael C Bassik, Lacramioara Bintu
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引用次数: 0

Abstract

Despite growing knowledge of the functions of individual human transcriptional effector domains, much less is understood about how multiple effector domains within the same protein combine to regulate gene expression. Here, we measure transcriptional activity for 8,400 effector domain combinations by recruiting them to reporter genes in human cells. In our assay, weak and moderate activation domains synergize to drive strong gene expression, whereas combining strong activators often results in weaker activation. In contrast, repressors combine linearly and produce full gene silencing, and repressor domains often overpower activation domains. We use this information to build a synthetic transcription factor whose function can be tuned between repression and activation independent of recruitment to target genes by using a small-molecule drug. Altogether, we outline the basic principles of how effector domains combine to regulate gene expression and demonstrate their value in building precise and flexible synthetic biology tools. A record of this paper's transparent peer review process is included in the supplemental information.

Abstract Image

转录激活因子和阻遏因子组合的高通量功能表征。
尽管人们对单个人类转录效应结构域的功能越来越了解,但对同一蛋白质内多个效应结构域如何结合调节基因表达的了解却少得多。在这里,我们通过招募8400个效应结构域组合到人类细胞中的报告基因来测量它们的转录活性。在我们的分析中,弱激活结构域和中等激活结构域协同驱动强基因表达,而结合强激活剂通常导致较弱的激活。相反,阻遏物线性结合并产生完全的基因沉默,并且阻遏物结构域通常压倒激活结构域。我们利用这些信息构建了一种合成转录因子,通过使用小分子药物,其功能可以在抑制和激活之间进行调节,而不依赖于靶基因的募集。总之,我们概述了效应域如何结合调节基因表达的基本原理,并证明了它们在构建精确灵活的合成生物学工具方面的价值。本文的透明同行评审过程记录包含在补充信息中。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cell Systems
Cell Systems Medicine-Pathology and Forensic Medicine
CiteScore
16.50
自引率
1.10%
发文量
84
审稿时长
42 days
期刊介绍: In 2015, Cell Systems was founded as a platform within Cell Press to showcase innovative research in systems biology. Our primary goal is to investigate complex biological phenomena that cannot be simply explained by basic mathematical principles. While the physical sciences have long successfully tackled such challenges, we have discovered that our most impactful publications often employ quantitative, inference-based methodologies borrowed from the fields of physics, engineering, mathematics, and computer science. We are committed to providing a home for elegant research that addresses fundamental questions in systems biology.
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