Orthogonality and Burdens of Heterologous AND Gate Gene Circuits in E. coli.,ACS Synthetic Biology

当前位置： X-MOL 学术 › ACS Synth. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Orthogonality and Burdens of Heterologous AND Gate Gene Circuits in E. coli.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-01-05 , DOI: 10.1021/acssynbio.7b00328
Qijun Liu _{1,

2,

3} , Jörg Schumacher ₄ , Xinyi Wan _{1,

2} , Chunbo Lou ₅ , Baojun Wang _{1,

2}

Affiliation

Synthetic biology approaches commonly introduce heterologous gene networks into a host to predictably program cells, with the expectation of the synthetic network being orthogonal to the host background. However, introduced circuits may interfere with the host's physiology, either indirectly by posing a metabolic burden and/or through unintended direct interactions between parts of the circuit with those of the host, affecting functionality. Here we used RNA-Seq transcriptome analysis to quantify the interactions between a representative heterologous AND gate circuit and the host Escherichia coli under various conditions including circuit designs and plasmid copy numbers. We show that the circuit plasmid copy number outweighs circuit composition for their effect on host gene expression with medium-copy number plasmid showing more prominent interference than its low-copy number counterpart. In contrast, the circuits have a stronger influence on the host growth with a metabolic load increasing with the copy number of the circuits. Notably, we show that variation of copy number, an increase from low to medium copy, caused different types of change observed in the behavior of components in the AND gate circuit leading to the unbalance of the two gate-inputs and thus counterintuitive output attenuation. The study demonstrates the circuit plasmid copy number is a key factor that can dramatically affect the orthogonality, burden and functionality of the heterologous circuits in the host chassis. The results provide important guidance for future efforts to design orthogonal and robust gene circuits with minimal unwanted interaction and burden to their host.

中文翻译：

大肠杆菌中异源和Gate基因回路的正交性和负担。

合成生物学方法通常将异源基因网络引入宿主以可预测地编程细胞，同时期望合成网络与宿主背景正交。然而，引入的电路可能通过造成代谢负担和/或通过电路的各个部分与宿主的那些之间的意外的直接相互作用而间接地干扰宿主的生理，从而影响功能。在这里，我们使用RNA-Seq转录组分析来量化代表异源与门电路与宿主大肠杆菌在各种条件下的相互作用，包括电路设计和质粒拷贝数。我们显示电路质粒拷贝数超过其对宿主基因表达的影响的电路组成，其中中等拷贝数质粒显示出比其低拷贝数对应物更突出的干扰。相反，随着新陈代谢负荷随电路拷贝数的增加，这些电路对宿主的生长有更强的影响。值得注意的是，我们显示出副本数量的变化，即从低副本到中副本，导致与门电路中组件行为的不同类型的变化，导致两个门输入的不平衡，从而导致输出衰减不直观。研究表明，电路质粒拷贝数是一个关键因素，可以极大地影响主机架中异源电路的正交性，负担和功能。

更新日期：2018-01-05

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11