Redesign of an Escherichia coli Nissle treatment for phenylketonuria using insulated genomic landing pads and genetic circuits to reduce burden
大肠杆菌
质粒
生物
基因
重组DNA
遗传学
作者
Alexander J. Triassi,Brandon D. Fields,Catherine E. Monahan,Jillian Means,Yong‐Jin Park,Hamid Doosthosseini,Padmakumar Jai,Vincent M. Isabella,Christopher A. Voigt
出处
期刊:Cell systems [Elsevier] 日期:2023-06-01卷期号:14 (6): 512-524.e12被引量:6
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.