Utilising datasheets for the informed automated design and build of a synthetic metabolic pathway

BackgroundThe automation of modular cloning methodologies permits the assembly of many genetic designs. Utilising characterised biological parts aids in the design and redesign of genetic pathways. The characterisation information held on datasheets can be used to determine whether a biological part meets the design requirements. To manage the design of genetic pathways, researchers have turned to modelling-based computer aided design software tools.ResultAn automated workflow has been developed for the design and build of heterologous metabolic pathways. In addition, to demonstrate the powers of electronic datasheets we have developed software which can transfer part information from a datasheet to the Design of Experiment software JMP. To this end we were able to use Design of Experiment software to rationally design and test randomised samples from the design space of a lycopene pathway in E. coli. This pathway was optimised by individually modulating the promoter strength, RBS strength, and gene order targets.ConclusionThe use of standardised and characterised biological parts will empower a design-oriented synthetic biology for the forward engineering of heterologous expression systems. A Design of Experiment approach streamlines the design-build-test cycle to achieve optimised solutions in biodesign. Developed automated workflows provide effective transfer of information between characterised information (in the form of datasheets) and DoE software.

[1]  S. Aves,et al.  Design of Experiments Methodology to Build a Multifactorial Statistical Model Describing the Metabolic Interactions of Alcohol Dehydrogenase Isozymes in the Ethanol Biosynthetic Pathway of the Yeast Saccharomyces cerevisiae. , 2018, ACS synthetic biology.

[2]  Christopher A. Voigt,et al.  Iterative algorithm-guided design of massive strain libraries, applied to itaconic acid production in yeast. , 2018, Metabolic engineering.

[3]  Carl-Fredrik Mandenius,et al.  Bioprocess optimization using design‐of‐experiments methodology , 2008, Biotechnology progress.

[4]  Erika Check Hayden,et al.  Synthetic biologists seek standards for nascent field , 2015, Nature.

[5]  Christopher A. Voigt,et al.  Principles of genetic circuit design , 2014, Nature Methods.

[6]  R. I. Kitney,et al.  Rapid Prototyping Platform for Saccharomyces cerevisiae Using Computer-Aided Genetic Design Enabled by Parallel Software and Workcell Platform Development , 2018, SLAS technology.

[7]  Tom Ellis,et al.  Synthetic Biology — A Primer:Revised Edition , 2015 .

[8]  J. Keasling,et al.  Engineering a mevalonate pathway in Escherichia coli for production of terpenoids , 2003, Nature Biotechnology.

[9]  Richard I. Kitney,et al.  Data model for biopart datasheets , 2017 .

[10]  Meghdad Hajimorad,et al.  BglBrick vectors and datasheets: A synthetic biology platform for gene expression , 2011, Journal of biological engineering.

[11]  M. De Mey,et al.  Standardization in synthetic biology: an engineering discipline coming of age , 2018, Critical reviews in biotechnology.

[12]  Zhen Zhang,et al.  Synthetic Biology Open Language (SBOL) Version 2.2.0 , 2018, J. Integr. Bioinform..

[13]  Huimin Zhao,et al.  Recent advances in DNA assembly technologies. , 2014, FEMS yeast research.

[14]  Christopher David Hirst,et al.  Automated BioPart characterisation for synthetic biology , 2014 .

[15]  Drew Endy,et al.  A survey of enabling technologies in synthetic biology , 2013, Journal of biological engineering.

[16]  Deepak Chandran,et al.  TinkerCell: modular CAD tool for synthetic biology , 2009, Journal of biological engineering.

[17]  Paul Freemont,et al.  The Foundry: the DNA synthesis and construction Foundry at Imperial College , 2016, Biochemical Society transactions.

[18]  Ya-bin Liu,et al.  Lycopene exerts anti-inflammatory effect to inhibit prostate cancer progression , 2018, Asian journal of andrology.

[19]  Iñaki Sainz de Murieta,et al.  Toward the First Data Acquisition Standard in Synthetic Biology. , 2016, ACS synthetic biology.

[20]  Huimin Zhao,et al.  Engineering biological systems using automated biofoundries. , 2017, Metabolic engineering.

[21]  Gary D. Bader,et al.  Promoting Coordinated Development of Community-Based Information Standards for Modeling in Biology: The COMBINE Initiative , 2015, Front. Bioeng. Biotechnol..

[22]  Tom Ellis,et al.  BASIC: A New Biopart Assembly Standard for Idempotent Cloning Provides Accurate, Single-Tier DNA Assembly for Synthetic Biology. , 2015, ACS synthetic biology.

[23]  Xinghuan Wang,et al.  Lycopene and Risk of Prostate Cancer , 2015, Medicine.

[24]  T. Ellis,et al.  Bricks and blueprints: methods and standards for DNA assembly , 2015, Nature Reviews Molecular Cell Biology.

[25]  Seon-Won Kim,et al.  Synthetic Biology and Metabolic Engineering for Marine Carotenoids: New Opportunities and Future Prospects , 2014, Marine drugs.

[26]  Carole Goble,et al.  An automated Design-Build-Test-Learn pipeline for enhanced microbial production of fine chemicals , 2018, Communications Biology.

[27]  Paulina Kanigowska,et al.  Smart DNA Fabrication Using Sound Waves , 2016, Journal of laboratory automation.

[28]  Drew Endy,et al.  Measuring the activity of BioBrick promoters using an in vivo reference standard , 2009, Journal of biological engineering.

[29]  Han N. Lim,et al.  Fundamental relationship between operon organization and gene expression , 2011, Proceedings of the National Academy of Sciences.

[30]  Adam Arkin,et al.  Setting the standard in synthetic biology , 2008, Nature Biotechnology.

[31]  D. Endy,et al.  Refinement and standardization of synthetic biological parts and devices , 2008, Nature Biotechnology.

[32]  W. R. Farmer,et al.  Improving lycopene production in Escherichia coli by engineering metabolic control , 2000, Nature Biotechnology.

[33]  Douglas Densmore,et al.  Design Automation in Synthetic Biology. , 2017, Cold Spring Harbor perspectives in biology.

[34]  Jacob Beal,et al.  Reducing DNA context dependence in bacterial promoters , 2017, PloS one.

[35]  Verena Siewers,et al.  Editorial: Yeast synthetic biology: new tools to unlock cellular function. , 2015, FEMS yeast research.

[36]  Erika Check Hayden Synthetic biology called to order. , 2015 .

[37]  Chris J. Myers,et al.  Tablet—next generation sequence assembly visualization , 2009, Bioinform..

[38]  G. Stephanopoulos,et al.  Improving Metabolic Pathway Efficiency by Statistical Model-Based Multivariate Regulatory Metabolic Engineering. , 2017, ACS synthetic biology.