Synthetic Biology for Everyday Life Our Project F ats ils rease - PowerPoint PPT Presentation

“Synthetic Biology for Everyday Life”

Our Project F ats ils rease s O G INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT Hayton Wastewater Treatment Facility 14 tonne fatberg found in Kingston, FINAL DESIGN & London, UK RESULTS SUMMARY 150,000 major blockages caused by FOGs every year in the UK. ATTRIBUTIONS

Hence… INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

SocioBricks INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES a background and basis for the project work: from social research to MODELLING Framework outreach PRODUCT to ensure projects are conducted ethically under legislation in Ethics different countries FINAL DESIGN & RESULTS strategies to accomplish the work's goals: be they answering a social Methods question to public engagement SUMMARY quantitative or qualitative, relating back to the initial framework and Analysis the methods employed to analyse the findings ATTRIBUTIONS the communication of the work done can be conveyed in several forms. In this collection, the user can find examples effective outputs Outputs from previous iGEM teams

SocioBricks INTRODUCTION Framework Ethics Methods Output SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

SocioBricks INTRODUCTION Framework Methods Analysis Output SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES Sheffield 2010 Sheffield 2014 MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Problem: Fatbergs INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Initial Design INTRODUCTION SOCIOBRICKS FAT Fat INITIAL DESIGN Breakdown POLICY & PRACTICES MODELLING PRODUCT Keratin FINAL DESIGN & Hair RESULTS Breakdown SUMMARY ATTRIBUTIONS

Initial Design INTRODUCTION SOCIOBRICKS INITIAL DESIGN Criteria POLICY & • Secrete enzyme to degrade fat  TliA PRACTICES lipase via type I secretion MODELLING • Secrete enzyme to degrade hair  PRODUCT KerUS Keratinase (self-secretion) FINAL DESIGN & RESULTS • Respond to fat levels  P fadR promoter SUMMARY ATTRIBUTIONS

Initial Design Criteria INTRODUCTION • Secrete lipid responsive lipase and keratinase enzymes SOCIOBRICKS LIPID RESPONSIVE LIPASE (BBa_K1498001) INITIAL DESIGN POLICY & PRACTICES Criteria P fadR RBS tliA Lipase Terminators BBa_K861060 BBa_K258006 MODELLING • Secrete lipid responsive lipase and TYPE I SECRETION MODULE (BBa_K258008) PRODUCT keratinase enzymes P tetR RBS PrtD Domain Terminator RBS PrtE Domain Terminator RBS PrtF Domain Terminators FINAL DESIGN & BBa_R0040 B0034 K258100 B0010 B0034 K258101 B0010 B0034 K258102 B0010, B0012 RESULTS LIPID RESPONSIVE KERATINASE SUMMARY ATTRIBUTIONS P fadR RBS kerUS Terminators BBa_K661060 (BBa_K1498000)

FadR Characterisation Bba_K861061 INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS TECAN data showing fluorescence produced from the fat responsive promoter / constitutive promoter over a 16 hour time period.

FadR Characterisation Bba_K861061 INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS TECAN data showing fluorescence produced from the fat responsive promoter / constitutive promoter over a 16 hour time period. SUMMARY • 4 different concentration of oleic acid ATTRIBUTIONS • No significant response to oleic acid by the promoter

FadR Characterisation • Cells DAPI blue stained INTRODUCTION to show placement SOCIOBRICKS • Grown in 2% Oleic Acid INITIAL DESIGN for 24 hours Constitutive RFP • RFP excited using 584nm POLICY & PRACTICES wavelength • Intensity of red linked to MODELLING amount of transcription PRODUCT fadR promoter- RFP from the promoter FINAL DESIGN & • No fluorescence seen in RESULTS P fadR cells – indicates no SUMMARY transcription from the fadR promoter promoter ATTRIBUTIONS Promoter shows no response to fat levels

Expert Consultation Framework Method Analysis Output Output INTRODUCTION SOCIOBRICKS Why? INITIAL DESIGN Understand urban drainage and the water industry • POLICY & PRACTICES Who? MODELLING Two leading academics working with water industry • PRODUCT What? FINAL DESIGN & RESULTS Implications for product design • SUMMARY – Compatible with wastewater treatment processes – Consider GMO release issues ATTRIBUTIONS

FOG Problems: Who’s to blame? INTRODUCTION Ethics Ethics Methods Analysis Output Ethics Framework SOCIOBRICKS INITIAL DESIGN Reason for study POLICY & PRACTICES • Develop insight from first consultations into the webs of responsibility relating to MODELLING FOG management PRODUCT FINAL DESIGN & RESULTS Methodology • Semi-structured interviews and visits SUMMARY designed to find out about differing views ATTRIBUTIONS on notions of responsibility

FOG Problems: Methodology INTRODUCTION SOCIOBRICKS INITIAL DESIGN Key Stakeholders POLICY & 6 Restaurant & Homeowner • PRACTICES • 3 Water Industry MODELLING 1 Council • PRODUCT FINAL DESIGN & RESULTS FOG problems at SUMMARY Hayton wastewater treatment facility ATTRIBUTIONS

FOG Problems: Findings INTRODUCTION SOCIOBRICKS INITIAL DESIGN Findings POLICY & PRACTICES Wide ranging practices • MODELLING Law on side of water industry • PRODUCT Consumer ratings • • Limitations FINAL DESIGN & RESULTS Private company Who’s to blame? • collecting used SUMMARY cooking oil Complex web of ATTRIBUTIONS practices

Nested Manufacturing INTRODUCTION The unit itself SOCIOBRICKS The bacteria in the unit INITIAL DESIGN The lipase from the bacteria POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Product Development INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES Analysis Output Framework Method Method MODELLING PRODUCT Personal Expert 8 Concepts Pro’s and Finished consultation con’s design research FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

In-Sewer Concept INTRODUCTION Larger scale installation that is SOCIOBRICKS easier to enforce and upkeep INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT Reduces sewer capacity FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Grease Trap Concept INTRODUCTION Already in use in restaurants and SOCIOBRICKS hence can modify existing units INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT Doesn’t protect from FINAL DESIGN & RESULTS detergents and is too BIG SUMMARY ATTRIBUTIONS

Perfusion Bioreactor INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT Too EXPENSIVE FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Vertical Stirred Tank Concept INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES Low oxygen diffusion MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

The Fatberglar INTRODUCTION SOCIOBRICKS INITIAL DESIGN POLICY & PRACTICES MODELLING PRODUCT FINAL DESIGN & RESULTS SUMMARY ATTRIBUTIONS

Summary INTRODUCTION SOCIOBRICKS Cheap to buy less than £100 INITIAL DESIGN Cheap to run • 132ml of water per day • bleach once a month POLICY & PRACTICES • freeze dried bacteria MODELLING • one nutrient block per month PRODUCT Easy to maintain 30 days between refills FINAL DESIGN & RESULTS Small enough for the kitchen sink SUMMARY ATTRIBUTIONS

Final Design Criteria INTRODUCTION • Secrete low dose lipase and keratinase SOCIOBRICKS enzymes continuously INITIAL DESIGN CONSTITUTIVE LIPASE (BBa_K1498001) POLICY & PRACTICES MODELLING Constitutive promoter RBS tliA Lipase Terminators BBa_J23110 BBa_K258006 PRODUCT TYPE I SECRETION MODULE (BBa_K258008) FINAL DESIGN & P tetR RBS PrtD Domain Terminator RBS PrtE Domain Terminator RBS PrtF Domain Terminators RESULTS BBa_R0040 B0034 K258100 B0010 B0034 K258101 B0010 B0034 K258102 B0010, B0012 SUMMARY CONSTITUTIVE KERATINASE (BBa_K1498000) ATTRIBUTIONS Constitutive promoter RBS kerUS Keratinase Terminators BBa_J23110 Bba_K1498000

Results: Lipase Assay for production of fatty acids by INTRODUCTION breakdown of lipids SOCIOBRICKS INITIAL DESIGN Empty POLICY & Plasmid & PRACTICES Exporter MODELLING PRODUCT Promoter, FINAL DESIGN & RESULTS Lipase & Exporter SUMMARY ATTRIBUTIONS 10 µl of cell culture supernatant on Rhodamine B plate Proof that our lipase works and is exported