Engineering compact bacterial combinatorial promoters for two-input chemical AND switching.
4 affiliations
- 1Jaramillo Lab, Warwick Integrative Synthetic Biology Centre and School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
- 2Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
- 3Jaramillo Lab, Warwick Integrative Synthetic Biology Centre and School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK. [email protected].
- 4Jaramillo Lab, i2SysBio, CSIC-University of Valencia, Paterna, Spain. [email protected].
Abstract
Engineering compact bacterial promoters that integrate two chemical inputs would simplify biosensors, synthetic circuits, and dynamic metabolic control, but such promoters remain difficult to build because a second operator can increase partial-state leak, reduce the fully induced state, and create sequence- or reporter-context effects. Here we engineered 12 compact Marionette-derived two-input promoter architectures in Escherichia coli by combining defined template scaffolds with added operators, then quantified 19 reporter-specific four-state truth tables under a predefined operational criterion for chemical-input AND behaviour. Single-input template-promoter controls in matched reporter contexts helped separate weak parent-scaffold output from effects introduced by the second operator. Nine curated architectures passed in at least one reporter, and selected constructs delivered high-utility AND responses, including [11]/max-off separation up to > 1,367. The full pass-fail dataset showed that a statistically higher [11] state does not necessarily create a useful switch. Practical utility depended mainly on suppressing [10] and [01] leakage, matching the template scaffold to the inserted operator, and controlling long-operator orientation and local sequence context. Reciprocal architectures behaved differently, and 4 of 7 dual-reporter architectures changed operational classification with reporter context. Thus, this work delivers a compact set of candidate Marionette-compatible two-input promoter parts and a source-traceable design framework for engineering bacterial chemical-input AND switches.
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BibTeX
@article{prakash2026engineering,
title = {{Engineering compact bacterial combinatorial promoters for two-input chemical AND switching.}},
author = {Satya Prakash and Alfonso Jaramillo},
journal = {Journal of biological engineering},
year = {2026},
month = jul,
doi = {10.1186/s13036-026-00728-8},
url = {https://europepmc.org/article/MED/42919391},
note = {CC BY},
}
RIS
TY - JOUR
TI - Engineering compact bacterial combinatorial promoters for two-input chemical AND switching.
AU - Satya Prakash
AU - Alfonso Jaramillo
PY - 2026
DA - 2026/07/01
JO - Journal of biological engineering
DO - 10.1186/s13036-026-00728-8
UR - https://europepmc.org/article/MED/42919391
AB - Engineering compact bacterial promoters that integrate two chemical inputs would simplify biosensors, synthetic circuits, and dynamic metabolic control, but such promoters remain difficult to build because a second operator can increase partial-state leak, reduce the fully induced state, and create sequence- or reporter-context effects. Here we engineered 12 compact Marionette-derived two-input promoter architectures in Escherichia coli by combining defined template scaffolds with added operators, then quantified 19 reporter-specific four-state truth tables under a predefined operational criterion for chemical-input AND behaviour. Single-input template-promoter controls in matched reporter contexts helped separate weak parent-scaffold output from effects introduced by the second operator. Nine curated architectures passed in at least one reporter, and selected constructs delivered high-utility AND responses, including [11]/max-off separation up to > 1,367. The full pass-fail dataset showed that a statistically higher [11] state does not necessarily create a useful switch. Practical utility depended mainly on suppressing [10] and [01] leakage, matching the template scaffold to the inserted operator, and controlling long-operator orientation and local sequence context. Reciprocal architectures behaved differently, and 4 of 7 dual-reporter architectures changed operational classification with reporter context. Thus, this work delivers a compact set of candidate Marionette-compatible two-input promoter parts and a source-traceable design framework for engineering bacterial chemical-input AND switches.
SN - 1754-1611
ER -