Feedforward growth rate control mitigates gene activation burden
<jats:title>Abstract</jats:title><jats:p>Heterologous gene activation causes non-physiological burden on cellular resources that cells are unable to adjust to. Here, we introduce a feedforward controller that actuates growth rate upon activation of a gene of interest (GOI) to compe...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Springer Science and Business Media LLC
2023
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| Online Access: | https://hdl.handle.net/1721.1/150903 |
| _version_ | 1826213987005497344 |
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| author | Barajas, Carlos Huang, Hsin-Ho Gibson, Jesse Sandoval, Luis Del Vecchio, Domitilla |
| author2 | Massachusetts Institute of Technology. Department of Mechanical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Mechanical Engineering Barajas, Carlos Huang, Hsin-Ho Gibson, Jesse Sandoval, Luis Del Vecchio, Domitilla |
| author_sort | Barajas, Carlos |
| collection | MIT |
| description | <jats:title>Abstract</jats:title><jats:p>Heterologous gene activation causes non-physiological burden on cellular resources that cells are unable to adjust to. Here, we introduce a feedforward controller that actuates growth rate upon activation of a gene of interest (GOI) to compensate for such a burden. The controller achieves this by activating a modified SpoT enzyme (SpoTH) with sole hydrolysis activity, which lowers ppGpp level and thus increases growth rate. An inducible RelA+ expression cassette further allows to precisely set the basal level of ppGpp, and thus nominal growth rate, in any bacterial strain. Without the controller, activation of the GOI decreased growth rate by more than 50%. With the controller, we could activate the GOI to the same level without growth rate defect. A cell strain armed with the controller in co-culture enabled persistent population-level activation of a GOI, which could not be achieved by a strain devoid of the controller. The feedforward controller is a tunable, modular, and portable tool that allows dynamic gene activation without growth rate defects for bacterial synthetic biology applications.</jats:p> |
| first_indexed | 2024-09-23T15:57:59Z |
| format | Article |
| id | mit-1721.1/150903 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T15:57:59Z |
| publishDate | 2023 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1509032023-06-10T03:03:55Z Feedforward growth rate control mitigates gene activation burden Barajas, Carlos Huang, Hsin-Ho Gibson, Jesse Sandoval, Luis Del Vecchio, Domitilla Massachusetts Institute of Technology. Department of Mechanical Engineering <jats:title>Abstract</jats:title><jats:p>Heterologous gene activation causes non-physiological burden on cellular resources that cells are unable to adjust to. Here, we introduce a feedforward controller that actuates growth rate upon activation of a gene of interest (GOI) to compensate for such a burden. The controller achieves this by activating a modified SpoT enzyme (SpoTH) with sole hydrolysis activity, which lowers ppGpp level and thus increases growth rate. An inducible RelA+ expression cassette further allows to precisely set the basal level of ppGpp, and thus nominal growth rate, in any bacterial strain. Without the controller, activation of the GOI decreased growth rate by more than 50%. With the controller, we could activate the GOI to the same level without growth rate defect. A cell strain armed with the controller in co-culture enabled persistent population-level activation of a GOI, which could not be achieved by a strain devoid of the controller. The feedforward controller is a tunable, modular, and portable tool that allows dynamic gene activation without growth rate defects for bacterial synthetic biology applications.</jats:p> 2023-06-09T18:02:56Z 2023-06-09T18:02:56Z 2022 2023-06-09T17:59:57Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/150903 Barajas, Carlos, Huang, Hsin-Ho, Gibson, Jesse, Sandoval, Luis and Del Vecchio, Domitilla. 2022. "Feedforward growth rate control mitigates gene activation burden." Nature Communications, 13 (1). en 10.1038/S41467-022-34647-1 Nature Communications Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Nature |
| spellingShingle | Barajas, Carlos Huang, Hsin-Ho Gibson, Jesse Sandoval, Luis Del Vecchio, Domitilla Feedforward growth rate control mitigates gene activation burden |
| title | Feedforward growth rate control mitigates gene activation burden |
| title_full | Feedforward growth rate control mitigates gene activation burden |
| title_fullStr | Feedforward growth rate control mitigates gene activation burden |
| title_full_unstemmed | Feedforward growth rate control mitigates gene activation burden |
| title_short | Feedforward growth rate control mitigates gene activation burden |
| title_sort | feedforward growth rate control mitigates gene activation burden |
| url | https://hdl.handle.net/1721.1/150903 |
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