title: Orthogonal intercellular signaling for programmed spatial behavior.
abstract: Bidirectional intercellular signaling is an essential feature of multicellular organisms, and the engineering of complex biological systems will require multiple pathways for intercellular signaling with minimal crosstalk. Natural quorum-sensing systems provide components for cell communication, but their use is often constrained by signal crosstalk. We have established new orthogonal systems for cell-cell communication using acyl homoserine lactone signaling systems. Quantitative measurements in contexts of differing receiver protein expression allowed us to separate different types of crosstalk between 3-oxo-C6- and 3-oxo-C12-homoserine lactones, cognate receiver proteins, and DNA promoters. Mutating promoter sequences minimized interactions with heterologous receiver proteins. We used experimental data to parameterize a computational model for signal crosstalk and to estimate the effect of receiver protein levels on signal crosstalk. We used this model to predict optimal expression levels for receiver proteins, to create an effective two-channel cell communication device. Establishment of a novel spatial assay allowed measurement of interactions between geometrically constrained cell populations via these diffusible signals. We built relay devices capable of long-range signal propagation mediated by cycles of signal induction, communication and response by discrete cell populations. This work demonstrates the ability to systematically reduce crosstalk within intercellular signaling systems and to use these systems to engineer complex spatiotemporal patterning in cell populations.
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK Computational Science Laboratory, Microsoft Research, Cambridge, UK.
authors.forename: Paul K
authors.initials: PK
authors.lastname: Grant
authors.affiliation: Computational Science Laboratory, Microsoft Research, Cambridge, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Dalchau
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK Computational Science Laboratory, Microsoft Research, Cambridge, UK.
authors.forename: James R
authors.initials: JR
authors.lastname: Brown
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK Departamento de Genética Molecular y Microbiologia, Facultad de Cs. Biológicas, Universidad Católica de Chile, Santiago, Chile.
authors.forename: Fernan
authors.initials: F
authors.lastname: Federici
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Rudge
authors.affiliation: Computational Science Laboratory, Microsoft Research, Cambridge, UK.
authors.forename: Boyan
authors.initials: B
authors.lastname: Yordanov
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK.
authors.forename: Om
authors.initials: O
authors.lastname: Patange
authors.affiliation: Computational Science Laboratory, Microsoft Research, Cambridge, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Phillips
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge, UK jh295@cam.ac.uk.
authors.forename: Jim
authors.initials: J
authors.lastname: Haseloff
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D002450:Cell Communication / Q000235:genetics*; D006714:Homoserine / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D053038:Quorum Sensing / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics*; D049490:Systems Biology
chemical_list: C109860:N-(3-oxododecanoyl)homoserine lactone; C088386:homoserine lactone; D006714:Homoserine; D015107:4-Butyrolactone
keywords: modeling; quorum sensing; spatial patterning; synthetic biology
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