Construction of a broad-host-range Anderson promoter series and particulate methane monooxygenase promoter variants expand the methanotroph genetic toolbox

Bhat, Etash H.; Henard, Jessica M.; Lee, Spencer A.; McHalffey, Dustin; Ravulapati, Mahith S.; Rogers, Elle V.; Yu, Logan; Skiles, David; Henard, Calvin A.

doi:10.1016/j.synbio.2024.02.003

Synthetic and Systems Biotechnology

2024

DOI: 10.1016/j.synbio.2024.02.003

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Construction of a broad-host-range Anderson promoter series and particulate methane monooxygenase promoter variants expand the methanotroph genetic toolbox

Etash H. Bhat,

Jessica M. Henard,

Spencer A. Lee

et al.

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Cited by 3 publications

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Industrializing methanotrophs and other methylotrophic bacteria: from bioengineering to product recovery

Sauvageau,

Stein,

Arenas

et al. 2024

Current Opinion in Biotechnology

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Industrializing methanotrophs and other methylotrophic bacteria: from bioengineering to product recovery

Sauvageau,

Stein,

Arenas

et al. 2024

Current Opinion in Biotechnology

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Methane to bioproducts: unraveling the potential of methanotrophs for biomanufacturing

Tan,

Ratra,

Singer

et al. 2024

Current Opinion in Biotechnology

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A genetically encoded biosensor to monitor dynamic changes of c-di-GMP with high temporal resolution

Kaczmarczyk,

van Vliet,

Jakob

et al. 2024

Nat Commun

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Monitoring changes of signaling molecules and metabolites with high temporal resolution is key to understanding dynamic biological systems. Here, we use directed evolution to develop a genetically encoded ratiometric biosensor for c-di-GMP, a ubiquitous bacterial second messenger regulating important biological processes like motility, surface attachment, virulence and persistence. The resulting biosensor, cdGreen2, faithfully tracks c-di-GMP in single cells and with high temporal resolution over extended imaging times, making it possible to resolve regulatory networks driving bimodal developmental programs in different bacterial model organisms. We further adopt cdGreen2 as a simple tool for in vitro studies, facilitating high-throughput screens for compounds interfering with c-di-GMP signaling and biofilm formation. The sensitivity and versatility of cdGreen2 could help reveal c-di-GMP dynamics in a broad range of microorganisms with high temporal resolution. Its design principles could also serve as a blueprint for the development of similar, orthogonal biosensors for other signaling molecules, metabolites and antibiotics.

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Construction of a broad-host-range Anderson promoter series and particulate methane monooxygenase promoter variants expand the methanotroph genetic toolbox

Cited by 3 publications

References 37 publications

Industrializing methanotrophs and other methylotrophic bacteria: from bioengineering to product recovery

Industrializing methanotrophs and other methylotrophic bacteria: from bioengineering to product recovery

Methane to bioproducts: unraveling the potential of methanotrophs for biomanufacturing

A genetically encoded biosensor to monitor dynamic changes of c-di-GMP with high temporal resolution

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