Team iGEM Paris-Saclay GEMOTE Project (Gene Expression Modulated by - - PowerPoint PPT Presentation

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Team iGEM Paris-Saclay GEMOTE Project (Gene Expression Modulated by - - PowerPoint PPT Presentation

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Team iGEM Paris-Saclay GEMOTE Project (Gene Expression Modulated by Temperature) Students : Audrey, Benjamin, Jrmie, Julien, Lucille, Pierre, Saber, Thibault, Yohann Advisors : Solenne, Nguyen Instructors : Jean-Luc, Philippe, Sylvie Outline

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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

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2 Team Paris-Saclay

The team

I

Scientific Project

II

Applications

III

Conclusion

IV

Outline

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TEAM PARIS-SACLAY

Team Paris-Saclay 3

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4 Team Paris-Saclay

Paris-Sud University

Facts on Paris-Sud University :

  • Part of the Paris-Saclay campus project
  • Second largest campus in Europe
  • Multidisciplinary university

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Paris

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5 Team Paris-Saclay

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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Team Paris-Saclay 6 6 Team Paris-Saclay

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Jérémie : 3rd year in Physical-Chemistry Thibault : 4th year in Biomolecules Engineering Yohann : 2nd year in Information technologies Audrey : 4th year in Applied Microbiology Doriane : 3rd year in Physical-Chemistry Lucille : 3rd year in Biotechnologies

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Team Paris-Saclay 7 7 Team Paris-Saclay

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Benjamin : 3rd year in Physical-Chemistry Séverine : 3rd year in Biotechnologies William : 3rd year in Biotechnologies Geoffrey : 3rd year in Biotechnologies Saber : 3rd year in Biotechnologies Pierre : 3rd year in Biology Julien : 2nd year in Information technologies

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SLIDE 8

Team Paris-Saclay 8 8 Team Paris-Saclay

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Solenne : PhD Student Advisor Nguyen : PhD Student Advisor Instructors (from the left to the right) :

  • Sylvie
  • Jean-Luc
  • Philippe
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SLIDE 9

Scientific Adventure Discovering Synthetic Biology Being part of a human project Have Fun ! Working with students from

  • ther fields

9 Team Paris-Saclay

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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10 Team Paris-Saclay

How ? Get informations about synthetic biology Being known in the University Looking for sponsors Spread knowledge

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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11 Team Paris-Saclay

Different levels of communication :

One year projects Long-term projects

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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12 Team Paris-Saclay

  • Presentation during the IGM days
  • Exhibition
  • Festival of Science

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

One year projects

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13 Team Paris-Saclay

  • Interviews of various players in synthetic biology
  • Increase interactions with the public using online exchange platforms
  • Organize conferences and public debates about synthetic biology

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Long-term projects

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14 Team Paris-Saclay

« Orsay, notre ville » « Le Parisien » « MCE TV » « Le Point : Etudiants » « Melty » « Paris-Sud Magazine »

Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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SCIENTIFIC PROJECT

Team Paris-Saclay 15

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16 Team Paris-Saclay

“GEMOTE” (Gene Expression MOdulated by TEmperature)

Objective : A new tool allowing gene expression in a specific temperature range.

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

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RNA thermometer

17 Team Paris-Saclay

Definition: sequences located in the 5’ untranslated regions of messenger RNAs with structures that depend on the temperature Mechanism: BBa_K115017

Scientific Project

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

RNA thermometer

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Thermosensitive repressor

18 Team Paris-Saclay

Definition: The cI protein is a transcriptional repressor from the phage lambda Mechanism: Temperature below 42°C Temperature above 42°C BBa_K539521

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

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Reporter system

19 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

BBa_K274100 The lycopene biosynthesis pathway Lycopene = red colored pigment, an intermediate in the biosynthesis

  • f many carotenoids.

Its biosynthesis needs 3 enzymes : CrtE, CrtB and CrtI. CrtE CrtB CrtI

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Combination by Gibson assembly of the GEMOTE construction :

  • RNA thermometer (BBa_K115017)
  • cI repressor (BBa_K092995)
  • Reporter system (BBa_K274100)
  • Terminator (BBa_361048)
  • Plasmid (pSB1A2)

20 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

How it works :

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21 Team Paris-Saclay

Below 32°C  RNA thermometer prevents CI expression  red bacteria

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

How it works :

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22 Team Paris-Saclay

Between 32°C and 42°C  translated and active CI protein  colorless bacteria

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

How it works :

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23 Team Paris-Saclay

Above 42°C  denatured CI repressor  red bacteria

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

How it works :

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24 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

How it works :

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25 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

Control 1

Wild type repressor

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26 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

26 Team Paris-Saclay

RBS without secondary structure

Control 2

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27 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

27 Team Paris-Saclay

The Gibson Assembly results for GEMOTE construction :

#11 control

  • What we expected: Red < 32°C < white < 42°C < red
  • What we observed :
  • After transformation : white colonies and one red #11
  • At 42°C : the white are still white and #11 is still red

Clone 11

Hypothesis : Some colonies harbor the GEMOTE construction but the temperature boundaries are shifted

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SLIDE 28
  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

Temperature-dependant expression of lycopene:

White colonies Red colonies White colonies

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29 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific Project

Reality: clone 11

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APPLICATIONS

Team Paris-Saclay 30

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Team Paris-Saclay 31

A temperature monitoring system :

 Assist scientists in their everyday work  Visual indication of the temperature at which the bacteria were grown

Applications

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion
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32 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Applications

Replacing the lycopene operon by a gene encoding a toxin Temperature Prison

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33 Team Paris-Saclay

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Applications

Temperature prison utilities Preventing GMMs dissemination Guardrail for probiotic bacteria

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SLIDE 34

CONCLUSION

Team Paris-Saclay 34

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35 Team Paris-Saclay

Conclusion

  • I. The team
  • II. Scientific

project

  • III. Applications
  • IV. Conclusion

Scientific adventure Discovering synthetic biology Being part of a human project Have fun ! Working with students from

  • ther fields
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36 Team Paris-Saclay

Thanks !