重新布线天然的转录后全局调控因子，以实现设计的、多层次的遗传回路。

Rewiring native post-transcriptional global regulators to achieve designer, multi-layered genetic circuits.

机构信息

McKetta Department of Chemical Engineering, The University of Texas at Austin, Austin, TX, 78712, USA.

Department of Chemistry and Biochemistry, Baylor University, Waco, TX, 76706, USA.

出版信息

Nat Commun. 2024 Oct 9;15(1):8752. doi: 10.1038/s41467-024-52976-1.

DOI:10.1038/s41467-024-52976-1

PMID:39384772

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11479628/

Abstract

As synthetic biology expands, creating "drag-and-drop" regulatory tools that can achieve diverse regulatory outcomes are paramount. Herein, we develop a approach for engineering complex post-transcriptional control by rewiring the Carbon Storage Regulatory (Csr) Network of Escherichia coli. We co-opt native interactions of the Csr Network to establish post-transcriptional logic gates and achieve complex bacterial regulation. First, we rationally engineer RNA-protein interactions to create a genetic toolbox of 12 BUFFER Gates that achieves a 15-fold range of expression. Subsequently, we develop a Csr-regulated NOT Gate by integrating a cognate 5' UTR that is natively Csr-activated into our platform. We then deploy the BUFFER and NOT gates to build a bi-directional regulator, two input Boolean Logic gates OR, NOR, AND and NAND and a pulse-generating circuit. Last, we port our Csr-regulated BUFFER Gate into three industrially relevant bacteria simply by leveraging the conserved Csr Network in each species.

摘要

随着合成生物学的发展，创建能够实现多样化调控效果的“拖放式”调控工具至关重要。在此，我们通过重新布线大肠杆菌的碳储存调控（Csr）网络，开发了一种用于工程复杂的转录后调控的方法。我们利用 Csr 网络的天然相互作用来建立转录后逻辑门，并实现复杂的细菌调控。首先，我们合理设计 RNA-蛋白相互作用，创建了一个由 12 个 BUFFER 门组成的遗传工具包，实现了 15 倍的表达范围。随后，我们通过将天然 Csr 激活的同源 5'UTR 整合到我们的平台中，开发了一个 Csr 调控的 NOT 门。然后，我们使用 BUFFER 和 NOT 门构建了一个双向调节剂、两个输入布尔逻辑门（OR、NOR、AND 和 NAND）和一个脉冲生成电路。最后，我们只需利用每个物种中保守的 Csr 网络，就将我们的 Csr 调控的 BUFFER 门移植到三种工业相关的细菌中。