UNITS_Trieste iGEM 2011 Competition World Championship Jamboree - PowerPoint PPT Presentation

... an innovative synthetic biome in which bacteria and eukaryotic cells need each other to survive! UNITS_Trieste iGEM 2011 Competition World Championship Jamboree November 5-7, 2011 MIT, Cambridge, Massachusetts, USA

INTRODUCTION EXPERIMENTS CONCLUSIONS « The Team: UNITS_Trieste »

INTRODUCTION EXPERIMENTS CONCLUSIONS « The living together of unlike organisms » « SYMBIOSIS » “ the living together of unlike organisms “ (de Bary, 1879)

INTRODUCTION EXPERIMENTS CONCLUSIONS « What is a consortium? » « all for one – one for all »

INTRODUCTION EXPERIMENTS CONCLUSIONS « What is a consortium? » 1 2 3 « e.g. hormones and autoinducers » autoinducers

INTRODUCTION EXPERIMENTS CONCLUSIONS « Quorum sensing (QS) » Bioluminescence Biofilm Virulence N-acyl-homoserine lactone « Regulation of synthetic bacterial consortia » « Does QS work also with eukaryotes? » Parasitism Infection Mutualism ?

INTRODUCTION EXPERIMENTS CONCLUSIONS INTRODUCTION EXPERIMENTS CONCLUSIONS « SynBiome » « SynBiome »

INTRODUCTION EXPERIMENTS CONCLUSIONS INTRODUCTION EXPERIMENTS CONCLUSIONS « SynBiome » « SynBiome »

INTRODUCTION EXPERIMENTS CONCLUSIONS INTRODUCTION EXPERIMENTS CONCLUSIONS « SynBiome » « SynBiome »

INTRODUCTION EXPERIMENTS CONCLUSIONS « SynBiome » Communication Rewards Mutualism

mRFP pSB1K3 Pconst. LasI Strain A pKBC12 sensor pKBC12 sensor Ctrl-

mRFP pSB1K3 Pconst. LasI OC12 Sample A Stds TIC intensity (CPS) OC8

pSB1K3 GFP PLasI β -gluco. Const. LasR PromLasI TraI Colony E Ctrl - CVO26 CVO26 Ctrl - Colony H OC8 0,0001ml 0,1 ml 0,00001ml 0,001ml 0,01 ml 0,1 uM 0,5 uL Std. Equivalent culture volume

N-(3-oxo-octanoyl)-homoserine lactone OC8 HLA Quantification 1400 Calibration curve 1200 1500 Miller Units 1000 1000 Miller Units 500 0 0 0,01 0,1 0,5 1 10 50 800 [nM OC8 HLA] 600 400 200 0 0 0,01 ul 0,1 ul 1 ul 0,01 ml 0,1 ml organic extraction equivalent volume Culture equivalent volume

Strain A Strain A Ctrl - Strain B CVO26 CVO26 CVO26 Strain B LB Agar

3OC 12 Sample A 46 Stds Culture media TIC intensity (CPS) 3OC 8

INTRODUCTION EXPERIMENTS CONCLUSIONS « Our BEST BioBrick » BBa_K553023 BBa_K553020 BBa_K553021

INTRODUCTION EXPERIMENTS CONCLUSIONS « Rewards: ß-glucosidase » GLUCOSE 0,2 OD 630 0,18 0,16 CELLOBIOSE 0,14 0,12 0,1 0,08 0,06 0,04 0,02 0 OC12 ind Const. const. 3OC12 ind ctrl - Ctrl- β -gluco β -gluco beta-gluco. beta-gluco.

INTRODUCTION EXPERIMENTS CONCLUSIONS « Rewards: ß-lactamase » TraBox CMVmin EGFP Sol β -Lactamase IRES Ampicillin pIRES – EGFP β -lactamase TraBox – sBla – IRES – EGFP + OC8 TraBox – sBla – IRES – EGFP

β -lactamase Const. Expressed Const. Expressed Not Induced Not Induced Ind. OXO C8 Ind. OXO C8 21 Kd sBLA 21 Kd sBLA α -Tub 64 Kd α -Tub 64 Kd Secreted Fraction Cellular Fraction

β -lactamase - OC8 + OC8

INTRODUCTION EXPERIMENTS CONCLUSIONS « SynBiome »

INTRODUCTION EXPERIMENTS CONCLUSIONS « Does it work? » • The INTRA -kingdom communication • The INTER -kingdom communication • The rewards: sBLA and Glucose

INTRODUCTION EXPERIMENTS CONCLUSIONS « Potential applications » S T R E S S S S T T R R E E S S S S «Libraries and complex molecules screening»

INTRODUCTION EXPERIMENTS CONCLUSIONS « The foundations » Design and give a POC of our system Have fun and full enjoy this experience Approach the synthetic biology field Looking for ideas Making brainstorming Reading papers Do the best for the best results

INTRODUCTION EXPERIMENTS CONCLUSIONS « Beyond the lab » Human Practices Standardize Researchers Night the standard Science Cafè Wiki theme Collaborations

INTRODUCTION EXPERIMENTS CONCLUSIONS « SYNBIOME @ Miami » “I am pleased to tell you That our abstract has been accepted at the 4th ASM Conference on Cell-Cell Communication in Bacteria .” November 6-9, 2011 Hyatt Regency Miami, Miami, Florida

... an innovative synthetic biome in which bacteria and eukaryotic cells need each other to survive! Question Time…