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Feb 28, 2018;6 (2): 206-215.e6.

Design of Adjacent Transcriptional Regions to Tune Gene Expression and Facilitate Circuit Construction.

Fuqing Wu, Qi Zhang, Xiao Wang
Author Information
  1. Fuqing Wu: School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  2. Qi Zhang: School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  3. Xiao Wang: School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA. Electronic address: xiaowang@asu.edu.
PMID: 29428414 DOI: 10.1016/j.cels.2018.01.010

Abstract

Polycistronic architecture is common for synthetic gene circuits, however, it remains unknown how expression of one gene is affected by the presence of other genes/noncoding regions in the operon, termed adjacent transcriptional regions (ATR). Here, we constructed synthetic operons with a reporter gene flanked by different ATRs, and we found that ATRs with high GC content, small size, and low folding energy lead to high gene expression. Based on these results, we built a model of gene expression and generated a metric that takes into account ATRs. We used the metric to design and construct logic gates with low basal expression and high sensitivity and nonlinearity. Furthermore, we rationally designed synthetic 5'ATRs with different GC content and sizes to tune protein expression levels over a 300-fold range and used these to build synthetic toggle switches with varying basal expression and degrees of bistability. Our comprehensive model and gene expression metric could facilitate the future engineering of more complex synthetic gene circuits.

Keywords

adjacent transcriptional region circuit design gene expression synthetic gene networks

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Grants

  1. R01 GM106081/NIGMS NIH HHS

MeSH Term

Animals
Gene Expression
Gene Expression Regulation
Gene Regulatory Networks
Genetic Engineering
Humans
Models, Genetic
Operon
Synthetic Biology
Transcription, Genetic
Journal Article
Article has an altmetric score of 3

Word Cloud

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