The COMET toolkit for composing customizable genetic programs in mammalian cells
The COMET toolkit for composing customizable genetic programs in mammalian cells
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DOI：
10.1038/s41467-019-14147-5
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发表时间：
2020-02-07
影响因子：
16.6
通讯作者：
Leonard, Joshua N.
中科院分区：
文献类型：
--
作者：
Donahue, Patrick S.;Draut, Joseph W.;Leonard, Joshua N.
设计哺乳动物细胞来执行复杂的和可定制的遗传程序需要一个由多个正交的和特征良好的转录因子(TF)组成的工具包。为了满足这一需求，我们开发了可组合的哺乳动物转录元件(COMET)--一组转录因子和启动子，能够设计和调节基因表达，而据我们所知，这是以前所不可能的。Comet目前包括44个激活锌指转录因子和12个抑制锌指转录因子，以及83个同源启动子，它们组合在一个框架中，很容易容纳新的部分。这个系统可以调节超过三个数量级的基因表达，提供化学诱导的转铁蛋白活性控制，并实现单层布尔逻辑。我们还开发了一个数学模型，它提供了对彗星性能特征的机械性见解。总而言之，Comet使在哺乳动物细胞中设计和构建可定制的遗传程序成为可能。
Engineering mammalian cells to carry out sophisticated and customizable genetic programs requires a toolkit of multiple orthogonal and well-characterized transcription factors (TFs). To address this need, we develop the COmposable Mammalian Elements of Transcription (COMET)-an ensemble of TFs and promoters that enable the design and tuning of gene expression to an extent not, to the best of our knowledge, previously possible. COMET currently comprises 44 activating and 12 inhibitory zinc-finger TFs and 83 cognate promoters, combined in a framework that readily accommodates new parts. This system can tune gene expression over three orders of magnitude, provides chemically inducible control of TF activity, and enables single-layer Boolean logic. We also develop a mathematical model that provides mechanistic insights into COMET performance characteristics. Altogether, COMET enables the design and construction of customizable genetic programs in mammalian cells.