Final IONIS-iGem poster2015.igem.org/files/poster/IONIS_Paris.pdf · 2015. 10. 7. · iGEM and...
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Popularization of biology in Elementary and High Schools Our team has organized some in- troduction courses for elementa- ry schools. They overviewed no- tions on: - DNA, - Living organisms (using the tree of life) The interest of students moti- vated us to have them extract banana's DNA. Another meeting was scheduled for High School students on: - Synthetic biology and iGEM - Human genetics. «Explain your project in 1 minute» This year the main goal of the IONIS iGEM team was to popularize Synthetic Biology. So, we have challenged other teams: They had to create a one-minute video intro- ducing their projects in the simplest way. BactMan Adventures is an in- novative mobile application popularizing science through educative mini-games around the life of a bacterium. The mobile application is available on all Android mobile phones. With it, you can discover biolo- gy through several games. The ap - Fun and ludic games - Information about the iGEM and synthetic biology Aware of the need to communicate and share about synthetic biology, IONIS Paris team a new way to popularize life sciences. In this way, the team has taken the challenge to develop a project based on two approaches : The Bio-Console : a real synthetic biology game BactMan Adventures: a mobile application presenting biology, synthetic biology and safety using mini-games. Photoinducible liposome Toxin Opening of the liposome and leakage of the toxins Photopolymerizable phospholipid (DC8,9PC) Laser beam (514nm) Matrix lipid (DPPC) 0,3mm 16mm 12mm x y Bubble detection: - Contrast between bubble and environment - «Hough Circle» detection algorithm - Find and return position Game part - Location Position in the image Interaction between real and virtual worlds - Game engine - GUI (Game user interface) - Graphical engine - Physical engine MFCS (Fluigent) : system (0-345 mbar) - advanced feedback control with no mechanical part THE BIO-CONSOLE Engineered bacterium Dead bacterium VISUALIZATION GAME OVER MICROFLUIDIC SYSTEM LOADING & ACTIVATION FLUORESCENCE BIOLUMINESCENCE Toxins KILLSWITCH & BLEACHING DRAINING LIPOSOME VVD N CAP VVD N CAP YC155 YN155 VVD N CAP VVD N CAP YN155 YC155 YN155 YC155 VVD N CAP 460nm 460nm LIGHT ILLUMINATION CONFORMATIONAL CHANGE HOMODIMERIZATION OF VVD PHOTORECEPTORS RECONSTITUTION OF THE SPLIT-YFP YC155 Linker O2 CO2 + + + LIGHT FURIMAZINE FURIMAMIDE NanoLuc Luciferase TM Oxigen oxidation of organic molecule Release of a visible photon Double T7 terminator reverse ccdB reverse RBS reverse 3’ 5’ Double T7 terminator reverse Endolysin reverse RBS reverse Holin reverse RBS reverse 3’ 5’ Double T7 terminator reverse HokD reverse RBS reverse 3’ 5’ 0 200000 400000 600000 800000 1000000 1200000 1400000 1600000 BBa_K325909 BBa_K1159001 T7-NanoLuc RLU 0 200000 400000 600000 800000 1000000 1200000 1400000 1600000 0.005 mM 0.01 mM 0.1 mM 0.5 mM 1 mM RLU Concentra on of IPTG T7-NanoLuc 0 200000 400000 600000 800000 1000000 1200000 1400000 0,01% 0,10% 0,50% 1% 2% RLU % of arabinose BBa_K1159001 Blue light stimulation induces VVD homo-dimerization and split YFP assembly. Blue light stimulation of pDawn allows toxins expression (HokD, ccdB or holin/ - endolysin) and cell lysis. The following parts have been designed in reverse to replace the MCS. FLUORESCENCE BIOLUMINESCENCE GAME OVER KILL SWITCH OVERVIEW MICROFLUIDICS Making the interface between real and virtual worlds Design of T7 promoter coupled to NanoLuc and its characterization: comparison with two luciferase biobricks (BBa_K1159001 and BBa_K325909) from the registry. NanoLuc construct of iGEM Munich 2013 (BBa_K1159001) was further characterized percentage of arabinose. NanoLuc CHARACTERIZATION 5’ 3’ T7 promoter Nanoluc T7 terminator PDMS chip (μfactory) - channel size: 300μm width, 50μm height 5’ MCS pR promoter cI 3’ Fix K2 promoter I promoter 9 YF1 Fix J 470nm The engineered light-responsive system YF1/FixJ within pDawn induces gene expression upon light activation. THE BIOCONSOLE : - bacteria - sponge ADVENTURES VIEW MORE ON OUR WIKI T7 terminator T7 promoter RBS VVD YC155 RBS VVD YN155 3’ 5’ P. Charleton , L. Chesnais , J. Chesnel , N. Cornille , H. Cremaschi , M. Da Costa , G. Defrel , A. Louisy , G. Mintec , M. Mohmmed , P-L. Nech , P. Trébulle , M. Zerhouni , G. Mercy and S. Juillot . 5 1 2 2 2 2 2 2 2 3 3 2 2 4 6 Affiliations 1 : E-artsup, Paris, France. 2 : Sup’Biotech, Paris, France. 3 : Epita, Paris, France. 4 : Paris Descartes University, Paris, France. 5 : Pasteur Institute, Paris, France. 6 : University of Freiburg, Freiburg, Germany. References Bilwes, A. M., Dunlap, J. C., & Crane, B. R. (2007). «Conformational Switching in the Fungal Light Sensor Vivid», 36(May), 1054-1058. Chang-Deng Hu, T. K. K. (2003). «Simultaneous visualization of multiple protein interactions in living cells using multicolor fluorescence complementation analysis». Nature Biotechnology, 21(5), 539-545. Hooper K, Ph.D (2012). «Application of a smaller, brighter, more versatile luciferase: NanoLuc Luciferase Technology». Ohlendorf,R., Vidavski R. R., Eldar A., Moffat K. & Möglich A. (2012). «From Dusk till Dawn: One-Plasmid System for Light-Regulated Gene Expression». 534-542. Müller K. & Weber W. (2013). «Optogenetic tools for mammalian systems». Mole- cular BioSystems, 9(4), 596-608. Yavlovich A., Singh A.n Blumenthal R., Purl A. (2011). «A novel class of photo-trig- gerable liposomes containing DPPC:DC8,9PC as vehicle for delivery of doxorubcin to cells». Biochim Biophys Acta - Biomembr., 1808(1), 117-26. TM
Transcript of Final IONIS-iGem poster2015.igem.org/files/poster/IONIS_Paris.pdf · 2015. 10. 7. · iGEM and...
Popularization of biology in Elementary and High Schools
Our team has organized some in-troduction courses for elementa-ry schools. They overviewed no-tions on:- DNA, - Living organisms (using the tree of life)
The interest of students moti-vated us to have them extract banana's DNA.
Another meeting was scheduled for High School students on: - Synthetic biology and iGEM - Human genetics.
«Explain your project in 1 minute»
This year the main goal of the IONIS iGEM team was to popularize Synthetic Biology. So, we have challenged other teams: They had to create a one-minute video intro-ducing their projects in the simplest way.
BactMan Adventures is an in-novative mobile application popularizing science through educative mini-games around the life of a bacterium. The mobile application is available on all Android mobile phones. With it, you can discover biolo-gy through several games.
The ap - Fun and ludic games - Information about the iGEM and synthetic biology
Aware of the need to communicate and share about synthetic biology, IONIS Paris teama new way to popularize life sciences. In this way, the team has taken the challenge to develop a project based on two approaches :
The Bio-Console : a real synthetic biology game
BactMan Adventures: a mobile application presenting biology, synthetic biology and safety using mini-games.
Photoinducibleliposome
Toxin Opening of the liposomeand leakage of the toxins
Photopolymerizablephospholipid (DC8,9PC)
Laser beam(514nm)
Matrix lipid(DPPC)
0,3mm
16mm
12mm
x
y
Bubble detection:- Contrast between bubble and environment- «Hough Circle» detection algorithm- Find and return position
Game part- Location Position in the image Interaction between real and virtual worlds- Game engine - GUI (Game user interface) - Graphical engine - Physical engine
MFCS (Fluigent) :
system (0-345 mbar) - advanced feedback control with no mechanical part
T H E B I O - C O N S O L E
Engineered bacterium
Deadbacterium
VISUALIZATION GAME OVER
MICROFLUIDIC SYSTEM
LOADING &ACTIVATION
FLUORESCENCE
BIOLUMINESCENCE
Toxins
KILLSWITCH &BLEACHING
DRAINING
LIPOSOME
VVD
NCAP VVD
NCAP
YC155
YN15
5
VVD
NCAP
VVD
NCAP
YN15
5
YC155 YN15
5
YC155
VVD
NCAP
460nm 460nm
LIGHT ILLUMINATION CONFORMATIONALCHANGE
HOMODIMERIZATIONOF VVD PHOTORECEPTORS
RECONSTITUTION OFTHE SPLIT-YFP
YC155
LinkerO2 CO2+ + + LIGHT
FURIMAZINE FURIMAMIDE
NanoLucLuciferase
TM
Oxigen oxidation of organic molecule
Release of avisible photon
Double T7 terminator reverse ccdB reverse RBS reverse
3’ 5’
Double T7 terminator reverse
Endolysinreverse RBS reverse
Holinreverse RBS reverse
3’ 5’
Double T7 terminator reverse HokD reverse RBS reverse
3’ 5’
0
200000
400000
600000
800000
1000000
1200000
1400000
1600000
BBa_K325909 BBa_K1159001 T7-NanoLuc
RLU
0
200000
400000
600000
800000
1000000
1200000
1400000
1600000
0.005 mM 0.01 mM 0.1 mM 0.5 mM 1 mM
RLU
Concentra on of IPTG
T7-NanoLuc
0
200000
400000
600000
800000
1000000
1200000
1400000
0,01% 0,10% 0,50% 1% 2%
RLU
% of arabinose
BBa_K1159001
Blue light stimulation induces VVD homo-dimerization and split YFP assembly.
Blue light stimulation of pDawn allows toxins expression (HokD, ccdB or holin/-endolysin) and cell lysis.The following parts have been designed in reverse to replace the MCS.
FLUORESCENCE BIOLUMINESCENCE
GAME OVER
KILL SWITCH
OVERVIEW
MICROFLUIDICS Making the interface between real and virtual worlds
Design of T7 promoter coupled to NanoLuc and its characterization: comparison with two luciferase biobricks (BBa_K1159001 and BBa_K325909) from the registry.
NanoLuc construct of iGEM Munich 2013 (BBa_K1159001) was further characterized
percentage of arabinose.
NanoLuc CHARACTERIZATION
5’ 3’
T7 promoterNanoluc T7 terminator
PDMS chip (µfactory)- channel size: 300µm width, 50µm height
5’MCS
pR promotercI
3’
Fix K2 promoter I promoter9
YF1 Fix J
470nm
The engineered light-responsive system YF1/FixJ within pDawn induces gene expression upon light activation.
THE BIOCONSOLE :
- bacteria- sponge
ADVENTURES
VIEW MOREON OUR
WIKI
T7 terminatorT7 promoter
RBS VVD YC155 RBS VVD YN155
3’ 5’
P. Charleton , L. Chesnais , J. Chesnel , N. Cornille , H. Cremaschi , M. Da Costa , G. Defrel , A. Louisy , G. Mintec , M. Mohmmed , P-L. Nech , P. Trébulle , M. Zerhouni , G. Mercy and S. Juillot .51 2 2 2 2 2 2 2 3 3 2 24 6
A�liations 1 : E-artsup, Paris, France. 2 : Sup’Biotech, Paris, France. 3 : Epita, Paris, France.
4 : Paris Descartes University, Paris, France. 5 : Pasteur Institute, Paris, France. 6 : University of Freiburg, Freiburg, Germany.
References Bilwes, A. M., Dunlap, J. C., & Crane, B. R. (2007). «Conformational Switching in the Fungal Light Sensor Vivid», 36(May), 1054-1058.
Chang-Deng Hu, T. K. K. (2003). «Simultaneous visualization of multiple protein interactions in living cells using multicolor �uorescence complementation analysis». Nature Biotechnology, 21(5), 539-545.
Hooper K, Ph.D (2012). «Application of a smaller, brighter, more versatile luciferase: NanoLuc Luciferase Technology».
Ohlendorf,R., Vidavski R. R., Eldar A., Mo�at K. & Möglich A. (2012). «From Dusk till Dawn: One-Plasmid System for Light-Regulated Gene Expression». 534-542.
Müller K. & Weber W. (2013). «Optogenetic tools for mammalian systems». Mole-cular BioSystems, 9(4), 596-608.
Yavlovich A., Singh A.n Blumenthal R., Purl A. (2011). «A novel class of photo-trig-gerable liposomes containing DPPC:DC8,9PC as vehicle for delivery of doxorubcin to cells». Biochim Biophys Acta - Biomembr., 1808(1), 117-26.
TM
