Synthetic gene expression perturbation systems with rapid, tunable, single-gene specificity in yeast.
Synthetic gene expression perturbation systems with rapid, tunable, single-gene specificity in yeast.
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在酵母中具有快速、可调、单基因特异性的合成基因表达扰动系统。
DOI：
10.1093/nar/gks1313
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发表时间：
2013-02-01
影响因子：
14.9
通讯作者：
Noyes MB
中科院分区：
文献类型：
--
作者：
McIsaac RS;Oakes BL;Wang X;Dummit KA;Botstein D;Noyes MB
一种动态控制真核生物中单基因表达的通用方法，无脱靶效应，是分子和系统生物学家长期寻求的工具。我们设计了两个人工转录因子（atf），它们含有小鼠转录因子Zif268的Cys2His2锌指dna结合域（特异性为9 bp）或合理设计的四个锌指阵列（特异性为12 bp）。这些结构域与人雌激素受体和VP16激活结构域融合表达。ATFs可以快速诱导一个由合成启动子驱动的单一基因，以响应引入一种惰性激素，而不会产生可检测到的脱靶效应。在缺乏诱导剂的情况下，合成的启动子无活性，无法检测到被调控的基因产物。加入诱导剂后，转录本在15分钟内被诱导50倍。我们提出了这些atf的定量表征，并提供了使其实现简单的结构。这些新工具允许动态地阐明调控网络元件，我们用一个主要的代谢调节因子Gcn4p证明了这一点。
A general method for the dynamic control of single gene expression in eukaryotes, with no off-target effects, is a long-sought tool for molecular and systems biologists. We engineered two artificial transcription factors (ATFs) that contain Cys2His2 zinc-finger DNA-binding domains of either the mouse transcription factor Zif268 (9 bp of specificity) or a rationally designed array of four zinc fingers (12 bp of specificity). These domains were expressed as fusions to the human estrogen receptor and VP16 activation domain. The ATFs can rapidly induce a single gene driven by a synthetic promoter in response to introduction of an otherwise inert hormone with no detectable off-target effects. In the absence of inducer, the synthetic promoter is inactive and the regulated gene product is not detected. Following addition of inducer, transcripts are induced >50-fold within 15 min. We present a quantitative characterization of these ATFs and provide constructs for making their implementation straightforward. These new tools allow for the elucidation of regulatory network elements dynamically, which we demonstrate with a major metabolic regulator, Gcn4p.
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影响因子：
56.9
作者：
Boch, Jens;Scholze, Heidi;Bonas, Ulla
通讯作者：
Bonas, Ulla
DOI：
10.1073/pnas.110149297
发表时间：
2000-06-20
影响因子：
11.1
作者：
Joung, JK;Ramm, EI;Pabo, CO
通讯作者：
Pabo, CO
影响因子：
4.5
作者：
Alexander, Ross D.;Barrass, J. David;Beggs, Jean D.
通讯作者：
Beggs, Jean D.
影响因子：
14.8
作者：
Heckman, Karin L.;Pease, Larry R.
通讯作者：
Pease, Larry R.
影响因子：
16
作者：
Elemento, Olivier;Slonim, Noam;Tavazoie, Saeed
通讯作者：
Tavazoie, Saeed