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Engineering a dynamic, controllable infectivity switch in bacteriophage T7

View ORCID ProfileChutikarn Chitboonthavisuk, Chun Huai Luo, View ORCID ProfilePhil Huss, Mikayla Fernholz, View ORCID ProfileSrivatsan Raman
doi: https://doi.org/10.1101/2021.08.23.457391
Chutikarn Chitboonthavisuk
1Dept. of Biochemistry, Univ. of Wisconsin-Madison, Madison, WI, USA
2Dept. of Bacteriology, Univ. of Wisconsin-Madison, Madison, WI, USA
3Microbiology Doctoral Training Program, University of Wisconsin-Madison
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  • ORCID record for Chutikarn Chitboonthavisuk
Chun Huai Luo
1Dept. of Biochemistry, Univ. of Wisconsin-Madison, Madison, WI, USA
2Dept. of Bacteriology, Univ. of Wisconsin-Madison, Madison, WI, USA
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Phil Huss
1Dept. of Biochemistry, Univ. of Wisconsin-Madison, Madison, WI, USA
2Dept. of Bacteriology, Univ. of Wisconsin-Madison, Madison, WI, USA
3Microbiology Doctoral Training Program, University of Wisconsin-Madison
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Mikayla Fernholz
1Dept. of Biochemistry, Univ. of Wisconsin-Madison, Madison, WI, USA
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Srivatsan Raman
1Dept. of Biochemistry, Univ. of Wisconsin-Madison, Madison, WI, USA
2Dept. of Bacteriology, Univ. of Wisconsin-Madison, Madison, WI, USA
4Dept. of Chemical & Biological Eng., Univ. of Wisconsin-Madison, Madison, WI, USA
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  • For correspondence: sraman4@wisc.edu
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Abstract

Transcriptional repressors play an important role in regulating phage genomes. Here, we examined how synthetic regulation based on repressors can be to create a dynamic, controllable infectivity switch in bacteriophage T7. We engineered T7 by replacing a large region of the early phage genome with combinations of ligand-responsive promoters and ribosome binding sites (RBS) designed to control the phage RNA polymerase. Phages with the engineered switch showed virulence comparable to wildtype when not repressed, indicating the phage can be engineered without a loss of fitness. When repressed, the most effective switch used a TetR promoter and a weak RBS, resulting in a two-fold increase in latent period (time to lyse host) and change in phage titer. Further, phage activity could be tuned by varying inducer concentrations. Our study provides a proof of concept for a simple circuit for user control over phage infectivity.

Competing Interest Statement

S.R. is on the scientific advisory board of MAP/PATH LLC. All other authors declare no competing financial interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted August 24, 2021.
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Engineering a dynamic, controllable infectivity switch in bacteriophage T7
Chutikarn Chitboonthavisuk, Chun Huai Luo, Phil Huss, Mikayla Fernholz, Srivatsan Raman
bioRxiv 2021.08.23.457391; doi: https://doi.org/10.1101/2021.08.23.457391
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Engineering a dynamic, controllable infectivity switch in bacteriophage T7
Chutikarn Chitboonthavisuk, Chun Huai Luo, Phil Huss, Mikayla Fernholz, Srivatsan Raman
bioRxiv 2021.08.23.457391; doi: https://doi.org/10.1101/2021.08.23.457391

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