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Team Chem

Team Chem. Biological Water Filtration. Purification Process. Add plants to possibly contaminated water Plant cells take in arsenic and store it Cells provide an output (color/light) signaling the water is clean or needs further processing Separate plants from water

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Team Chem

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  1. Team Chem Biological Water Filtration

  2. Purification Process • Add plants to possibly contaminated water • Plant cells take in arsenic and store it • Cells provide an output (color/light) signaling the water is clean or needs further processing • Separate plants from water • Possibility for further processing

  3. Operating System

  4. Operating System

  5. Devices • 2 Arsenic Sensors • activated by any arsenic (2.5 ppb) • activated by maximum tolerable arsenic level • 2 Light Generators • Arsenic Transport and Storage Device

  6. Arsenic Transport and Storage Arsenic Measurement Remove the gene that encodes this transport protein Tripathi, R.D., Srivastava, S., Mishra, S., Singh, N., Tuli, R., Gupta, D.K. and Maathuis, F. Arsenic Hazards: Strategies for Tolerance and Remediation by Plants. Trends in Biotechnology. 2007, 25.4, 158-165.

  7. 2.5 ppb Arsenic Sensor Arsenic Transport and Storage Device (constitutively active) Max Arsenic Sensor Device Level Diagram Blue Light Generator Red Light Generator

  8. 2.5 ppb Arsenic Sensor Arsenic Transport and Storage Device (constitutively active) Max Arsenic Sensor Simulation: Pure Water Blue Light Generator Red Light Generator Result: No Light → Separation

  9. 2.5 ppb [As] Sensor As As Arsenic Transport and Storage Device (constitutively active) MAXIMUM ARSENIC Max Arsenic Sensor Simulation: Maximum Arsenic Blue Light Generator Red Light Generator Result: Cells Emit Red Light → Separate and Process Again

  10. 1 1 1 1 1 0 0 0 0 0 t=0 No Arsenic Timing Diagram: Maximum Arsenic Blue Light Generator 2.5 ppb [As] Sensor Transport and Storage Maximum [As] Sensor Red Light Generator Contaminated Water Added Midlevel [As] Maximum [As]

  11. 1 1 1 1 1 0 0 0 0 0 t=0 No Arsenic Timing Diagram: Midlevel Arsenic Blue Light Generator 2.5 ppb [As] Sensor Transport and Storage Maximum [As] Sensor Red Light Generator Contaminated Water Added Midlevel [As] Maximum [As]

  12. Biological PartsBackground System ARR2 ARR1 Arsenic resistance genes reduce arsenic so it can be complexed to the storage protein

  13. Biological Parts: Genetic Circuit Maximum P(Y) Constitutively active AtPCS1 AtABBC1 AtABBC2 Arsenic Transport P(ArsR) P(X) BLGD BBa_K325909 2.5 ppb Arsenic P(ArsD) RLGD BBa_K325219 Maximum Arsenic X-Repressor Y-Repressor Constitutively active ArsR BBa_J33201 ArsD Max Arsenic Arsenic Detectors 2.5 ppb Arsenic

  14. Biological Parts: Genetic Circuit P(Y) Constitutively active AtPCS1 AtABBC1 AtABBC2 Arsenic Transport P(ArsR) P(X) BLGD BBa_K325909 2.5 ppb Arsenic P(ArsD) RLGD BBa_K325219 Maximum Arsenic X-Repressor Y-Repressor Constitutively active ArsR BBa_J33201 ArsD Max Arsenic Arsenic Detectors 2.5 ppb Maximum Result: Blue Light Result: Red Light Arsenic

  15. Parts List

  16. Cellular Chassis [Source:http://www.botany.hawaii.edu/nlc_biology/1406/lab/r2/slide6.jpg] [Source: http://www.ppws.vt.edu/scott/weed_id/eldde.htm]

  17. Nucleus vs. Chloroplasts • Chloroplasts • Genes cannot be passed on by pollination • Very difficult to control how genes are passed to new generations • Nucleus • Cross-pollination would be a major issue • Crops that take in and store arsenic • Solution: • Elodea can be asexual • Easy to implant genes – a plant can grow from a few cells • No threat of cross-breeding

  18. Testing/Debugging [Source: http://www.sciencephoto.com/images/] • Cell Death • Effectiveness of Transport and Storage of Arsenic • Need to determine vacuole limits - maximum tolerable arsenic concentration • Testing Light Generators • Promoter Design • ArsR works for concentrations 2.5 ppb • ArsD must be tuned to experimentally determined maximum arsenic concentration

  19. Global Impacts

  20. Impacts If this process succeeds: • Solves one of the most problematic issues of the developing world, saving millions of lives. • 70 million people are affected in Bangladesh alone - arsenic in ground water is the cause of 23% of all the deaths there!

  21. Kanchan Arsenic Filter • $35 per filter • 90% efficient (brings down to 50 ppb if there was 500 initially). • Returns back to the ground at the end.

  22. Why Biological Water Filtration by Team Chemistry? • No energy requirement (operates independent of electricity) • Accessibility to effective water filtration technology even in rural parts of the developing world • Very low cost of sustaining system • The end product is not a waste • Plants could be sent to central processing center when full. Arsenic could be removed and used for chemotherapy. • More efficient than any other existing arsenic filters.

  23. Open Issues • The cost of entire project is unknown. • Legal issues attached to introducing genetically modified organisms into the environment? • Length of effectiveness is unknown • (How long until the organism reaches max arsenic uptake capacity?) • The rate of arsenic uptake is unknown. • The process of genetically modifying plants is very slow.

  24. Citations • Bobrowicz, P et al. “Isolation of three contiguous genes, ACR1, ACR2 and ACR3, involved in resistance to arsenic compounds in the yeast Saccharomyces cerevisiae.” Yeast (Chichester, England) 13.9 (1997): 819-28. <http://www.ncbi.nlm.nih.gov/pubmed/9234670>. • Deepesh N. De.Plant cell vacuoles: an introduction. CSIRO Publishing, Collingwood, 2000. • Dhingra, Amit, and Henry Daniell. “Chloroplast genetic engineering via organogenesis or somatic embryogenesis.” Methods in molecular biology (Clifton, N.J.) 323.6 (2006): 245-62. http://www.ncbi.nlm.nih.gov/pubmed/16892222.  • Song, W.-Y. et al. “Arsenic tolerance in Arabidopsis is mediated by two ABCC-type phytochelatin transporters.” Proceedings of the National Academy of Sciences 18 (2010). 17 Nov. 2010 <http://www.pnas.org/cgi/doi/10.1073/pnas.1013964107>. • Tripathi, Rudra D et al. “Arsenic hazards: strategies for tolerance and remediation by plants.” Trends in biotechnology 25.4 (2007): 158-65. 9 Jul. 2010 <http://www.ncbi.nlm.nih.gov/pubmed/17306392>.

  25. Citations Kanchan Water Filter: http://web.mit.edu/watsan/tech_hwts_chemical_kanchanarsenicfilter.html State of Washington, Department of Ecology: http://www.ecy.wa.gov/programs/wq/plants/weeds/aqua002.html Registry of Standard Biological Parts: http://partsregistry.org/Main_Page National Center for Biotechnology Information: http://www.ncbi.nlm.nih.gov/ Sacchromyces Genome Database:www.yeastgenome.org The Oligator (to remove unwanted restriction sites): http://gcat.davidson.edu/igem10/index.html Translation Map (Sequence Manipulation Suite- to make sure mutations didn’t change the proteins): http://biosyn.avxtrk.net/Gizmo/Tools/SMS/trans_map.html

  26. Acknowledgements Dr. June Medford, Department of Biology, Colorado State University Susan Murcott, Department of Civil and Environmental Engineering, MIT Dr. Chris French, Institute of Structural and Molecular Biology, School of Biological Sciences, University of Edinburgh Dr. Natalie Kuldell 20.385 Mentors: Yuan, Tina, Andrew and Pei.

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