Splash presentation tra slides
- 1. Unveiling the mysteries of synthetic biology with Presenters: Jonlin Chen, Grace Livermore, Tim Abbott, and Jeffrey Ly
- 2. WHAT IS SYNTHETIC BIOLOGY? Designing and constructing biological devices, networks and pathways for useful purposes Creating synthetic components or re-assembling pre-existing genes Wiring biological circuitry to make: Biological switches, oscillators, toggle switches Logic gates, pulse generators For: Biosensing, therapeutic treatment, biofuels…. And much more
- 3. WHAT IS IGEM? • ‘International Genetically Engineered Machines” • Global synthetic biology competition • HS / Undergraduate / Graduate / Entrepreneurship • Students design and build complex biological systems made from standard, interchangeable DNA parts “Biobricks”
- 4. How does one genetically modify an organism? Herbert Boyer Robert Swanson
- 5. • What is a restriction enzyme? DNA Restriction Enzyme • Why do bacteria have these enzymes? • Immunity against viruses! • Will the enzymes cut up the bacteria’s DNA? • Nope! The bacteria is able to protect its own DNA!
- 6. What Made These Two Interested in Restriction Enzymes? Herbert Boyer Robert Swanson • What if you could cut open bacterial DNA, and insert DNA of your choice in?
- 7. DNA DNA Recombinant DNA
- 8. History of iGEM • Started as a spin-off of an MIT course in 2003 • Has grown from 5 teams in 2004 to 245 in 2013! • Competition is run by co-founder: Randy Rettberg • Established the BioBrick, a standardized biological part for easier genetic manipulation
- 9. How do You Clone with BioBricks? 1. Get some DNA! 5 mL culture Plasmid DNA E. coli
- 10. How do You Clone with BioBricks? 2. Amplify target sequence through PCR
- 11. How do You Clone with BioBricks? 3. Double-digest backbone and PCR product
- 12. How do You Clone with BioBricks? 4. Gel purify your digests
- 13. How do You Clone with BioBricks? 5. De-phosphorylate backbone
- 14. How do You Clone with BioBricks? 6. Ligate your DNA
- 15. How do You Clone with BioBricks? 7. Transform into bacteria
- 16. The Team ● Award-winning undergraduate project team ● International Genetically Engineered Machines ● “Biobricks” : Standardized genetic parts ○ Ever-expanding registry ● Annual competition
- 17. Presenting Cornell iGem’s 2014 Project…
- 18. “Lead It Go!”
- 19. “LEAD IT GO” 2014 HEAVY METAL SEQUESTRATION SYSTEM • Engineering E. coli for the uptake of heavy metals • Integrate cells into dry lab’s filtration system • Test efficacy of filter with lab based and local water supplies • Building CAD portfolio of designs that allow the system to scale to many applications
- 20. Water Contamination ● Heavy metals o Mercury, lead, nickel ● Sources: o Industrial manufacturing o Hydraulic fracturing o Coal mining o Construction ● Harmful to humans, ecosystem, aquatic life o Biomagnification
- 21. Project Goals ● Engineer E.coli to uptake heavy metals from the environment ● Integrate cells into a continuous water filtration system ● Test efficacy of biological filter with lab based and local water samples
- 22. Heavy Metal Transport Proteins ● Proteins specific to heavy metals o nixA - nickel o merT and merP - mercury o CBP4 - lead ● Behind the constitutive Anderson promoter nixA Anderson Promoter
- 23. E.coli - Metal Transport E.coli Metal Ions Transport Protein
- 24. Metallothioneins ● Low molecular weight proteins ● High metal binding affinities o GST-YMT, GST-PMT ● Expression causes cellular dormancy ● Arabinose inducible system Arabinose araBAD Promoter Metallothionein T7 Promoter T7 Polymerase
- 25. Metallothionein Expression E.coli Metal Ions Transport Protein Metallothioneins
- 26. Metallothionein Expression E.coli Metal Ions Transport Protein Metallothioneins
- 27. Saturation Sensing System ● Insert mRFP gene behind metal-inducible promoters o Cells will fluoresce when metallothioneins are saturated ● Metal sensitive promoters induced once MT are saturated
- 28. Reporter System E.coli Metal Ions Transport Protein Metallothioneins Reporter Plasmid
- 29. Efficacy Analysis ● Continuous flow hollow fiber reactor ● Implementation o Factory waste water outlet o Contaminated bodies of water ● Testing o Lab made waste water o Local water samples ▪ Cayuga Lake ▪ Ithaca Gun factory
- 30. Fun Activity: Learn to Load Gels! Purpose: To determine relative lengths of pieces of DNA Applications of DNA Electrophoresis ● murder mysteries ● paternity cases ● disease diagnosis
- 31. Examples of Successful Gels
- 32. Thank you for coming! ● Special thanks to o Cornell University o College of Engineering o Wilson Lab o Dr. Shivaun Archer o Corning