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UNITS_Trieste
iGEM 2011 Competition World Championship Jamboree
... an innovative synthetic biome in which bacteria and eukaryotic cells need each other to survive!
November 5-7, 2011 MIT, Cambridge, Massachusetts, USA
UNITS_Trieste iGEM 2011 Competition World Championship Jamboree - - PowerPoint PPT Presentation
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
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« SYMBIOSIS »
“ the living together of unlike organisms “ (de Bary, 1879)
« The living together of unlike organisms »
INTRODUCTION EXPERIMENTS CONCLUSIONS
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« all for one – one for all »
« What is a consortium? »
INTRODUCTION EXPERIMENTS CONCLUSIONS
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1 2 3
« e.g. hormones and autoinducers » autoinducers
« What is a consortium? »
INTRODUCTION EXPERIMENTS CONCLUSIONS
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N-acyl-homoserine lactone Bioluminescence
Biofilm
Virulence
« Regulation of synthetic bacterial consortia » « Does QS work also with eukaryotes? »
Parasitism Infection Mutualism ?
« Quorum sensing (QS) »
INTRODUCTION EXPERIMENTS CONCLUSIONS
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« SynBiome »
INTRODUCTION EXPERIMENTS CONCLUSIONS
« SynBiome»
INTRODUCTION EXPERIMENTS CONCLUSIONS
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« SynBiome »
INTRODUCTION EXPERIMENTS CONCLUSIONS
« SynBiome»
INTRODUCTION EXPERIMENTS CONCLUSIONS
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« SynBiome »
INTRODUCTION EXPERIMENTS CONCLUSIONS
« SynBiome»
INTRODUCTION EXPERIMENTS CONCLUSIONS
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Communication Rewards Mutualism
« SynBiome »
INTRODUCTION EXPERIMENTS CONCLUSIONS
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Ctrl-
pKBC12 sensor Strain A pKBC12 sensor
pSB1K3
- Pconst. LasI
mRFP
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pSB1K3
- Pconst. LasI
mRFP
TIC intensity (CPS) Sample A Stds OC12 OC8
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0,0001ml 0,1 ml 0,01 ml 0,001ml Std. 0,1 uM 0,5 uL OC8
Equivalent culture volume
0,00001ml
Colony E Colony H CVO26 CVO26 Ctrl - Ctrl -
pSB1K3
β-gluco. PLasI GFP
- Const. LasR
PromLasI TraI
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200 400 600 800 1000 1200 1400 0,01 ul 0,1 ul 1 ul 0,01 ml 0,1 ml
Miller Units
- rganic extraction equivalent volume
OC8 HLA Quantification
500 1000 1500 0,01 0,1 0,5 1 10 50 Miller Units [nM OC8 HLA]
Calibration curve Culture equivalent volume
N-(3-oxo-octanoyl)-homoserine lactone
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CVO26 CVO26 Strain A Ctrl - Strain A Strain B
LB Agar
CVO26
Strain B
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3OC 8 3OC 12 Sample A 46 Stds Culture media TIC intensity (CPS)
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« Our BEST BioBrick »
INTRODUCTION EXPERIMENTS CONCLUSIONS BBa_K553023 BBa_K553020 BBa_K553021
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CELLOBIOSE GLUCOSE
« Rewards: ß-glucosidase »
INTRODUCTION EXPERIMENTS CONCLUSIONS
0,02 0,04 0,06 0,08 0,1 0,12 0,14 0,16 0,18 0,2
const. beta-gluco. 3OC12 ind beta-gluco. ctrl -
OD630
Const. β-gluco OC12 ind β-gluco Ctrl-
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« Rewards: ß-lactamase »
INTRODUCTION EXPERIMENTS CONCLUSIONS
TraBox CMVmin Sol β-Lactamase EGFP IRES pIRES – EGFP TraBox – sBla – IRES – EGFP + OC8 TraBox – sBla – IRES – EGFP
Ampicillin β -lactamase
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Secreted Fraction Cellular Fraction
64 Kd
- Const. Expressed
- Ind. OXO C8
Not Induced 21 Kd α-Tub sBLA 64 Kd 21 Kd α-Tub sBLA
- Const. Expressed
- Ind. OXO C8
Not Induced
β -lactamase
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- OC8
+ OC8 β -lactamase
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INTRODUCTION EXPERIMENTS CONCLUSIONS
« SynBiome »
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INTRODUCTION EXPERIMENTS CONCLUSIONS
« Does it work? »
- The INTRA-kingdom communication
- The INTER-kingdom communication
- The rewards: sBLA and Glucose
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INTRODUCTION EXPERIMENTS CONCLUSIONS
«Libraries and complex molecules screening»
« Potential applications »
S T R E S S S T R E S S S T R E S S
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Design and give a POC of our system
«The foundations»
INTRODUCTION EXPERIMENTS CONCLUSIONS
Approach the synthetic biology field
Have fun and full enjoy this experience
Looking for ideas
Reading papers
Making brainstorming
Do the best for the best results
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« Beyond the lab »
Collaborations INTRODUCTION EXPERIMENTS CONCLUSIONS
Researchers Night Science Cafè Wiki theme Standardize the standard
Human Practices
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« SYNBIOME @ Miami »
INTRODUCTION EXPERIMENTS CONCLUSIONS “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
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