Team iGEM Paris-Saclay GEMOTE Project (Gene Expression Modulated by Temperature) Students : Audrey, Benjamin, Jérémie, Julien, Lucille, Pierre, Saber, Thibault, Yohann Advisors : Solenne, Nguyen Instructors : Jean-Luc, Philippe, Sylvie
Outline The team I Scientific Project II Applications III Conclusion IV Team Paris-Saclay 2
TEAM PARIS-SACLAY Team Paris-Saclay 3
Team Paris-Saclay Paris I. The team II. Scientific project Paris-Sud University Facts on Paris-Sud University : III. Applications - Part of the Paris-Saclay campus project - Second largest campus in Europe - Multidisciplinary university IV. Conclusion Team Paris-Saclay 4
Team Paris-Saclay I. The team II. Scientific project III. Applications IV. Conclusion Team Paris-Saclay 5
Team Paris-Saclay Jérémie : 3 rd year in Physical-Chemistry Thibault : I. The team 4 th year in Biomolecules Engineering II. Scientific project Audrey : 4th year in Applied Microbiology Yohann : III. Applications 2 nd year in Information technologies IV. Conclusion Doriane : 3 rd year in Lucille : 3 rd year in Physical-Chemistry Biotechnologies Team Paris-Saclay Team Paris-Saclay 6 6
Team Paris-Saclay Julien : 2 nd year in Saber : Information technologies 3 rd year in Biotechnologies I. The team Séverine : II. Scientific 3 rd year in project Biotechnologies III. Applications Pierre : 3 rd year in Benjamin : Biology 3 rd year in IV. Conclusion Physical-Chemistry Geoffrey : 3 rd year in William : 3 rd year in Biotechnologies Biotechnologies Team Paris-Saclay Team Paris-Saclay 7 7
Team Paris-Saclay Solenne : PhD Student Advisor I. The team II. Scientific project Nguyen : III. Applications PhD Student Advisor IV. Conclusion Instructors (from the left to the right) : - Sylvie - Jean-Luc - Philippe Team Paris-Saclay Team Paris-Saclay 8 8
Team Paris-Saclay Scientific Adventure I. The team Working with Discovering II. Scientific project students from Synthetic other fields Biology III. Applications IV. Conclusion Being part of a Have Fun ! human project Team Paris-Saclay 9
Team Paris-Saclay Get informations about How ? I. The team synthetic biology II. Scientific project Being known in Looking for the University sponsors III. Applications IV. Conclusion Spread knowledge Team Paris-Saclay 10
Team Paris-Saclay Different levels of communication : I. The team II. Scientific project III. Applications One year projects IV. Conclusion Long-term projects Team Paris-Saclay 11
Team Paris-Saclay One year projects I. The team • Presentation during the IGM days II. Scientific project • Exhibition III. Applications • Festival of Science IV. Conclusion Team Paris-Saclay 12
Team Paris-Saclay Long-term projects I. The team • Interviews of various players in synthetic biology II. Scientific project • Increase interactions with the public using online exchange platforms III. Applications • Organize conferences and public debates about synthetic biology IV. Conclusion Team Paris-Saclay 13
Team Paris-Saclay « Orsay, notre ville » « Le Parisien » I. The team « MCE TV » II. Scientific project « Melty » III. Applications « Le Point : Etudiants » IV. Conclusion « Paris-Sud Magazine » Team Paris-Saclay 14
SCIENTIFIC PROJECT Team Paris-Saclay 15
Scientific Project I. The team Objective : A new tool allowing gene expression in a specific temperature range. II. Scientific project III. Applications IV. Conclusion “GEMOTE” (Gene Expression MOdulated by TEmperature) Team Paris-Saclay 16
Scientific Project RNA thermometer I. The team Definition: sequences located in the 5’ untranslated regions of messenger RNAs with structures that depend on the temperature II. Scientific project Mechanism: III. Applications IV. Conclusion RNA thermometer BBa_K115017 Team Paris-Saclay 17
Scientific Project Thermosensitive repressor I. The team Definition: The cI protein is a transcriptional repressor from the phage lambda II. Scientific project Mechanism: III. Applications IV. Conclusion Temperature below 42°C Temperature above 42°C BBa_K539521 Team Paris-Saclay 18
Scientific Project Reporter system I. The team Lycopene = red colored pigment, an intermediate in the biosynthesis of many carotenoids. II. Scientific project Its biosynthesis needs 3 enzymes : CrtE, CrtB and CrtI. The lycopene biosynthesis pathway III. Applications IV. Conclusion CrtE CrtB CrtI BBa_K274100 Team Paris-Saclay 19
Scientific Project How it works : I. The team II. Scientific project III. Applications Combination by Gibson assembly of the GEMOTE construction : • RNA thermometer (BBa_K115017) • cI repressor (BBa_K092995) IV. Conclusion • Reporter system (BBa_K274100) • Terminator (BBa_361048) • Plasmid (pSB1A2) Team Paris-Saclay 20
Scientific Project How it works : I. The team II. Scientific project III. Applications IV. Conclusion Below 32°C RNA thermometer prevents CI expression red bacteria Team Paris-Saclay 21
Scientific Project How it works : I. The team II. Scientific project III. Applications IV. Conclusion Between 32°C and 42°C translated and active CI protein colorless bacteria Team Paris-Saclay 22
Scientific Project How it works : I. The team II. Scientific project III. Applications IV. Conclusion Above 42°C denatured CI repressor red bacteria Team Paris-Saclay 23
Scientific Project How it works : I. The team II. Scientific project III. Applications IV. Conclusion Team Paris-Saclay 24
Scientific Project Wild type repressor Control 1 I. The team II. Scientific project III. Applications IV. Conclusion Team Paris-Saclay 25
Scientific Project Control 2 RBS without secondary structure I. The team II. Scientific project III. Applications IV. Conclusion Team Paris-Saclay Team Paris-Saclay 26 26
Scientific Project The Gibson Assembly results for GEMOTE construction : • What we expected: Red < 32°C < white < 42°C < red • What we observed : I. The team • After transformation : white colonies and one red #11 • At 42°C : the white are still white and #11 is still red II. Scientific project III. Applications IV. Conclusion Clone 11 control #11 Hypothesis : Some colonies harbor the GEMOTE construction but the temperature boundaries are shifted Team Paris-Saclay Team Paris-Saclay 27 27
Scientific Project Temperature-dependant expression of lycopene: I. The team II. Scientific project White colonies Red colonies III. Applications IV. Conclusion White colonies
Scientific Project I. The team II. Scientific project III. Applications Reality: clone 11 IV. Conclusion Team Paris-Saclay 29
APPLICATIONS Team Paris-Saclay 30
Applications A temperature monitoring system : I. The team Assist scientists in their everyday work Visual indication of the temperature II. Scientific project at which the bacteria were grown III. Applications IV. Conclusion Team Paris-Saclay 31
Applications Replacing the lycopene operon by a gene encoding a toxin I. The team II. Scientific project III. Applications Temperature Prison IV. Conclusion Team Paris-Saclay 32
Applications Temperature I. The team prison utilities II. Scientific project III. Applications Preventing Guardrail for IV. Conclusion GMMs probiotic dissemination bacteria Team Paris-Saclay 33
CONCLUSION Team Paris-Saclay 34
Conclusion Scientific adventure I. The team Working with Discovering II. Scientific project students from synthetic other fields biology III. Applications IV. Conclusion Being part of a Have fun ! human project Team Paris-Saclay 35
Thanks ! Team Paris-Saclay 36
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