Programmable T7-based synthetic transcription factors

Nucleic Acids Research
Brendan J Hussey, David R McMillen

Abstract

Despite recent progress on synthetic transcription factor generation in eukaryotes, there remains a need for high-activity bacterial versions of these systems. In synthetic biology applications, it is useful for transcription factors to have two key features: they should be orthogonal (influencing only their own targets, with minimal off-target effects), and programmable (able to be directed to a wide range of user-specified transcriptional start sites). The RNA polymerase of the bacteriophage T7 has a number of appealing properties for synthetic biological designs: it can produce high transcription rates; it is a compact, single-subunit polymerase that has been functionally expressed in a variety of organisms; and its viral origin reduces the connection between its activity and that of its host's transcriptional machinery. We have created a system where a T7 RNA polymerase is recruited to transcriptional start sites by DNA binding proteins, either directly or bridged through protein-protein interactions, yielding a modular and programmable system for strong transcriptional activation of multiple orthogonal synthetic transcription factor variants in Escherichia coli. To our knowledge this is the first exogenous, programmable ac...Continue Reading

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Citations

Jan 30, 2019·Chembiochem : a European Journal of Chemical Biology·Jinyue PuBryan C Dickinson
Jan 30, 2020·Nature Reviews. Microbiology·M Fata Moradali, Bernd H A Rehm
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May 7, 2019·ACS Synthetic Biology·Abigail J SmithNigel J Savery
Oct 2, 2019·ACS Synthetic Biology·Leo Y T Chou, William M Shih
Jul 25, 2019·Current Opinion in Chemical Biology·William M DawsonDerek N Woolfson
Jul 7, 2021·Nature Communications·I Cody MacDonaldTara L Deans

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Methods Mentioned

BETA
two hybrid
restriction digest

Software Mentioned

ZiFit
CoDA
OPEN

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