Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, 60208, USA.
Interdisciplinary Biological Sciences Program, Northwestern University, Evanston, IL, 60208, USA.
Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
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.
工程化哺乳动物细胞以执行复杂且可定制的遗传程序需要一套多种正交且特征良好的转录因子（TF）工具。为满足这一需求，我们开发了 COmposable Mammalian Elements of Transcription（COMET）-一组 TF 和启动子，使基因表达的设计和调整达到了我们所知的前所未有的程度。COMET 目前包含 44 种激活和 12 种抑制锌指 TF 以及 83 种同源启动子，组合在一个易于容纳新部件的框架中。该系统可以在三个数量级上调整基因表达，提供化学诱导的 TF 活性控制，并实现单层布尔逻辑。我们还开发了一种数学模型，为 COMET 的性能特征提供了机制见解。总之，COMET 使哺乳动物细胞中可定制遗传程序的设计和构建成为可能。
