Redesign of an Escherichia coli Nissle treatment for phenylketonuria using insulated genomic landing pads and genetic circuits to reduce burden.

IF 9 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Cell Systems Pub Date : 2023-06-21 DOI:10.1016/j.cels.2023.05.004

Alexander J Triassi, Brandon D Fields, Catherine E Monahan, Jillian M Means, Yongjin Park, Hamid Doosthosseini, Jai P Padmakumar, Vincent M Isabella, Christopher A Voigt

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Abstract

To build therapeutic strains, Escherichia coli Nissle (EcN) have been engineered to express antibiotics, toxin-degrading enzymes, immunoregulators, and anti-cancer chemotherapies. For efficacy, the recombinant genes need to be highly expressed, but this imposes a burden on the cell, and plasmids are difficult to maintain in the body. To address these problems, we have developed landing pads in the EcN genome and genetic circuits to control therapeutic gene expression. These tools were applied to EcN SYNB1618, undergoing clinical trials as a phenylketonuria treatment. The pathway for converting phenylalanine to trans-cinnamic acid was moved to a landing pad under the control of a circuit that keeps the pathway off during storage. The resulting strain (EcN SYN8784) achieved higher activity than EcN SYNB1618, reaching levels near when the pathway is carried on a plasmid. This work demonstrates a simple system for engineering EcN that aids quantitative strain design for therapeutics.

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利用绝缘基因组着陆垫和基因电路重新设计治疗苯丙酮尿症的大肠杆菌尼氏疗法，以减轻负担。

为了构建治疗菌株，人们设计了大肠杆菌 Nissle（EcN）来表达抗生素、毒素降解酶、免疫调节剂和抗癌化疗药物。为了达到疗效，重组基因需要高度表达，但这给细胞带来了负担，而且质粒在体内难以维持。为了解决这些问题，我们在 EcN 基因组中开发了着陆垫和基因回路，以控制治疗基因的表达。这些工具已应用于正在进行苯丙酮尿症临床试验的 EcN SYNB1618。将苯丙氨酸转化为反式肉桂酸的途径被转移到了一个着陆垫上，该着陆垫由一个电路控制，可在储存期间关闭该途径。由此产生的菌株（EcN SYN8784）比 EcN SYNB1618 具有更高的活性，达到了质粒上携带该途径时的水平。这项工作展示了一种简单的 EcN 工程系统，有助于定量设计用于治疗的菌株。

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 Medicine-Pathology and Forensic Medicine

CiteScore

16.50

自引率

1.10%

发文量

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.