Antisense transcription as a tool to tune gene expression
Name
Brophy-2016-Antisense transcript.pdf
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1.15 MB
Format
Adobe PDF
Checksum (MD5)
de4617e635c64aee3e8045147ad88ae9
Author(s)
Brophy, Jennifer Ann
Voigt, Christopher A.
Date Issued
January 2016
Journal
Molecular Systems Biology
Publisher
Nature Publishing Group
Citation
Brophy, J. A., and C. A. Voigt. “Antisense Transcription as a Tool to Tune Gene Expression.” Molecular Systems Biology 12, no. 1 (January 14, 2016): 854–854.
Version
Final published version
Abstract
A surprise that has emerged from transcriptomics is the prevalence of genomic antisense transcription, which occurs counter to gene orientation. While frequent, the roles of antisense transcription in regulation are poorly understood. We built a synthetic system in Escherichia coli to study how antisense transcription can change the expression of a gene and tune the response characteristics of a regulatory circuit. We developed a new genetic part that consists of a unidirectional terminator followed by a constitutive antisense promoter and demonstrate that this part represses gene expression proportionally to the antisense promoter strength. Chip‐based oligo synthesis was applied to build a large library of 5,668 terminator–promoter combinations that was used to control the expression of three repressors (PhlF, SrpR, and TarA) in a simple genetic circuit (NOT gate). Using the library, we demonstrate that antisense promoters can be used to tune the threshold of a regulatory circuit without impacting other properties of its response function. Finally, we determined the relative contributions of antisense RNA and transcriptional interference to repressing gene expression and introduce a biophysical model to capture the impact of RNA polymerase collisions on gene repression. This work quantifies the role of antisense transcription in regulatory networks and introduces a new mode to control gene expression that has been previously overlooked in genetic engineering.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Massachusetts Institute of Technology. Department of Biological Engineering
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.15252/msb.20156540
