Orthogonal intercellular signaling for programmed spatial behavior

Grant, Paul K.; Dalchau, Neil; Brown, Jo; Federici, Fernán; Rudge, Tim; Yordanov, Boyan; Patange, Om; Phillips, Andrew; Haseloff, Jim

doi:10.15252/msb.20156590

Cited by 73 publications

(147 citation statements)

References 31 publications

(41 reference statements)

Supporting

Mentioning

140

Contrasting

Order By: Relevance

“…The use of consortia of organisms could potentially alleviate these bottlenecks if the functionality of the genetic circuit can be distributed among different cell populations 8 . Additionally, the use of synthetic consortia could expand the capabilities and applications of synthetic biology by enabling compartmentalisation 9 , cell specialisation 10 , parallel bio-computation 11 , increased bioprocess efficiency 12 and spatial organisation 13 .…”

mentioning

confidence: 99%

“…Various studies have sought to increase the number of available AHL communication modules with functional devices built from the biological components of the las 18 , tra 18 , rpa 18 , rhl 19 , cin 19 , and esa 20 quorum sensing systems. Nevertheless, quorum sensing devices can exhibit various degrees of crosstalk either in the form of AHL molecules binding to non-cognate transcription factors (chemical crosstalk) or transcription factors binding to non-cognate promoters (genetic cross-talk) 13 . The degree of orthogonality between designed AHL communication modules can be quantified by high-throughput screening as demonstrated for a set of designed AHL-receiver devices of the lux, las, rpa, and tra quorum systems 18 .…”

mentioning

confidence: 99%

“…For this set of devices only the tra and rpa devices were determined to be completely orthogonal for both chemical and genetic crosstalk 18 . However, a number of strategies have been demonstrated for minimising crosstalk, which include modulating the expression levels of the transcription factor that influence the response function of the device, and quorum sensing promoter engineering 13 .…”

mentioning

confidence: 99%

See 2 more Smart Citations

Tools for engineering coordinated system behaviour in synthetic microbial consortia

Kylilis

Stan

Polizzi

2017

Preprint

View full text Add to dashboard Cite

Advancing synthetic biology to the multicellular level requires the development of multiple orthogonal cell-to-cell communication channels to propagate information with minimal signal interference. The development of quorum sensing devices, the cornerstone technology for building microbial communities with coordinated system behaviour, has largely focused on reducing signal leakage between systems of cognate AHL/transcription factor pairs. However, the use of noncognate signals as a design feature has received limited attention so far. Here, we demonstrate the largest library of AHL-receiver devices constructed to date with all cognate and non-cognate chemical signal interactions quantified and we develop a software tool that allows automated selection of orthogonal chemical channels. We use this approach to identify up to four orthogonal channels in silico and experimentally demonstrate the simultaneous use of three channels in coculture. The development of multiple non-interfering cell-to-cell communication channels will facilitate the design of synthetic microbial consortia for novel applications including distributed biocomputation, increased bioprocess efficiency, cell specialisation, and spatial organisation.

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Tools for engineering coordinated system behaviour in synthetic microbial consortia

Kylilis

Stan

Polizzi

2017

Preprint

View full text Add to dashboard Cite

show abstract

“…To fulfill the design of mutual suppression of motility, CheZ is constitutively expressed from λ promoter (P λ ) that can be repressed by CI repressor. A NOT gate is implemented by placing CI under the control of P lux and P las81 , a mutated P lux with stronger binding affinity with LasR [30], such that high intracellular AHLs (reflecting high cell densities) activate CI expression and in turn shut off the cheZ transcription. The two inhibitors constitutively express fluorescent markers mCherry and sfGFP respectively.…”

Section: Methodsmentioning

confidence: 99%

Engineering cooperative patterns in multi-species bacterial colonies

Curatolo

Zhou²,

Zhao³

et al. 2019

Preprint

View full text Add to dashboard Cite

Self-organization is a hallmark of all living systems [1]. In particular, coordinated cellular behavior, commonly orchestrated at the population level through reciprocal interactions among different cell species [2][3][4], regulates the spatial arrangement of specialized cell types to generate tissue patterning and form complex body layouts [5,6]. The overwhelming complexity of living systems, however, makes deciphering the underlying mechanisms difficult and limits our knowledge of basic pattern-forming mechanism in vivo [7,8]. A successful strategy is then to work with synthetic, engineered systems, in which cellular interactions can be more easily tailored and studied [9][10][11][12][13]. Here, we demonstrate a simple mechanism through which different populations of cells can self-organize in periodic patterns. Programmed population interactions are shown to lead to coordinated out-ofphase spatial oscillations of two engineered populations of Escherichia coli. Using a combination of experimental and theoretical approaches, we show how such patterns arise autonomously from reciprocal density-dependent activation of cellular motility between the two species, without the need of any preexisting positional or orientational cues. Moreover, by re-designing the interaction, the original out-of-phase spatial oscillation rhythm of the two populations can be accordingly turned into in-phase oscillations. The robustness and versatility of the underlying pattern-formation process suggest that it could both be generically encountered in nature, for instance in the complex bacterial ecosystems found in biofilms [14][15][16], and used to promote the mixing or demixing of active particles in a controlled way.We engineered Escherichia coli AMB1655 into two genetically distinct strains so as to enhance the * These authors contributed equally † Corresponding authors:

show abstract

“…Specific gene regulatory programs are able to translate the spatiotemporal information provided by the local concentration of morphogen gradients into robust gene expression patterns (Wolpert, 1996, Green and Sharpe, 2015, Rogers and Schier, 2011. This mechanism has been extensively used in synthetic systems to generate spatial patterns, especially stripe patterns, which were produced through synthetic feed-forward loops (Basu et al, 2005, Schaerli et al, 2014, inducible promoters (Grant et al, 2016) and AND-gates (Boehm et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Controlling spatiotemporal pattern formation in a concentration gradient with a synthetic toggle switch

Barbier

Pérez-Carrasco

Schaerli

2019

Preprint

View full text Add to dashboard Cite

The formation of spatiotemporal patterns of gene expression is frequently guided by gradients of diffusible signaling molecules. The toggle switch subnetwork, composed of two cross-repressing transcription factors, is a common component of gene regulatory networks in charge of patterning, converting the continuous information provided by the gradient into discrete abutting stripes of gene expression. We present a synthetic biology framework to understand and characterize the spatiotemporal patterning properties of the toggle switch. To this end, we built a synthetic toggle switch controllable by diffusible molecules in Escherichia coli. We analyzed the patterning capabilities of the circuit by combining quantitative measurements with a mathematical reconstruction of the underlying dynamical system. The toggle switch can produce robust patterns with sharp boundaries, governed by bistability and hysteresis. We further demonstrate how the hysteresis, position, timing, and precision of the boundary can be controlled, highlighting the dynamical flexibility of the circuit.

show abstract

Orthogonal intercellular signaling for programmed spatial behavior

Cited by 73 publications

References 31 publications

Tools for engineering coordinated system behaviour in synthetic microbial consortia

Tools for engineering coordinated system behaviour in synthetic microbial consortia

Engineering cooperative patterns in multi-species bacterial colonies

Controlling spatiotemporal pattern formation in a concentration gradient with a synthetic toggle switch

Contact Info

Product

Resources

About