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Title: Jungle Express is a versatile repressor system for tight transcriptional control

Abstract

Tightly regulated promoters are essential for numerous biological applications, where strong inducibility, portability, and scalability are desirable. Current systems are often incompatible with large-scale fermentations due to high inducer costs and strict media requirements. Here, we describe the bottom-up engineering of ‘Jungle Express’, an expression system that enables efficient gene regulation in diverse proteobacteria. This system is guided by EilR, a multidrug-binding repressor with high affinity to its optimized operator and cationic dyes that act as powerful inducers at negligible costs. In E. coli, the engineered promoters exhibit minimal basal transcription and are inducible over four orders of magnitude by 1 µM crystal violet, reaching expression levels exceeding those of the strongest current bacterial systems. Further, we provide molecular insights into specific interactions of EilR with its operator and with two inducers. The versatility of Jungle Express opens the way for tightly controlled and efficient gene expression that is not restricted to host organism, substrate, or scale.

Authors:
; ; ; ; ; ; ; ORCiD logo; ORCiD logo; ORCiD logo; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Inst. of Health (NIH) (United States); Howard Hughes Medical Inst. (United States); Swiss National Science Foundation (SNSF)
OSTI Identifier:
1467933
Alternate Identifier(s):
OSTI ID: 1477413
Grant/Contract Number:  
AC02-05CH11231; AC52-07NA27344; SC3GM096943
Resource Type:
Journal Article: Published Article
Journal Name:
Nature Communications
Additional Journal Information:
Journal Name: Nature Communications Journal Volume: 9 Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; applied microbiology; synthetic biology; transcriptional regulatory elements

Citation Formats

Ruegg, Thomas L., Pereira, Jose H., Chen, Joseph C., DeGiovanni, Andy, Novichkov, Pavel, Mutalik, Vivek K., Tomaleri, Giovani P., Singer, Steven W., Hillson, Nathan J., Simmons, Blake A., Adams, Paul D., and Thelen, Michael P. Jungle Express is a versatile repressor system for tight transcriptional control. United Kingdom: N. p., 2018. Web. doi:10.1038/s41467-018-05857-3.
Ruegg, Thomas L., Pereira, Jose H., Chen, Joseph C., DeGiovanni, Andy, Novichkov, Pavel, Mutalik, Vivek K., Tomaleri, Giovani P., Singer, Steven W., Hillson, Nathan J., Simmons, Blake A., Adams, Paul D., & Thelen, Michael P. Jungle Express is a versatile repressor system for tight transcriptional control. United Kingdom. doi:10.1038/s41467-018-05857-3.
Ruegg, Thomas L., Pereira, Jose H., Chen, Joseph C., DeGiovanni, Andy, Novichkov, Pavel, Mutalik, Vivek K., Tomaleri, Giovani P., Singer, Steven W., Hillson, Nathan J., Simmons, Blake A., Adams, Paul D., and Thelen, Michael P. Thu . "Jungle Express is a versatile repressor system for tight transcriptional control". United Kingdom. doi:10.1038/s41467-018-05857-3.
@article{osti_1467933,
title = {Jungle Express is a versatile repressor system for tight transcriptional control},
author = {Ruegg, Thomas L. and Pereira, Jose H. and Chen, Joseph C. and DeGiovanni, Andy and Novichkov, Pavel and Mutalik, Vivek K. and Tomaleri, Giovani P. and Singer, Steven W. and Hillson, Nathan J. and Simmons, Blake A. and Adams, Paul D. and Thelen, Michael P.},
abstractNote = {Tightly regulated promoters are essential for numerous biological applications, where strong inducibility, portability, and scalability are desirable. Current systems are often incompatible with large-scale fermentations due to high inducer costs and strict media requirements. Here, we describe the bottom-up engineering of ‘Jungle Express’, an expression system that enables efficient gene regulation in diverse proteobacteria. This system is guided by EilR, a multidrug-binding repressor with high affinity to its optimized operator and cationic dyes that act as powerful inducers at negligible costs. In E. coli, the engineered promoters exhibit minimal basal transcription and are inducible over four orders of magnitude by 1 µM crystal violet, reaching expression levels exceeding those of the strongest current bacterial systems. Further, we provide molecular insights into specific interactions of EilR with its operator and with two inducers. The versatility of Jungle Express opens the way for tightly controlled and efficient gene expression that is not restricted to host organism, substrate, or scale.},
doi = {10.1038/s41467-018-05857-3},
journal = {Nature Communications},
issn = {2041-1723},
number = 1,
volume = 9,
place = {United Kingdom},
year = {2018},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1038/s41467-018-05857-3

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Cited by: 2 works
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