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Biodesign for the real world

Biodesign for the real world . Falco ENZLER and Bastien ORSET. General overview. What is Biodesign ? What is our kit and why make a new one? What about the legal aspects? An example of approved field kit: ARSOlux What about confinement? DNA construct details

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Biodesign for the real world

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  1. Biodesign for the real world Falco ENZLER and Bastien ORSET

  2. General overview • WhatisBiodesign? • Whatisour kit and whymake a new one? • What about the legal aspects? • An example of approvedfield kit: ARSOlux • What about confinement? • DNA constructdetails • Future Design

  3. Biodesign Project • Biodesign is a multidisciplinary project with different research aspects: • development of field compatible water quality sensing systems • portable real-time sensing • crowd-sourcedmapping This projectis a collaboration between people fromIndia, Indonesia and Switzerland • We are students in ourthirdyear of life science bachelorat the EPFL and wejoined the Biodesign’s team for ourbachelor’sproject. http://biodesign.cc/

  4. Whatisourproject? • The aim of thisprojectis to detect arsenic in water samples in the field. • This detectionisaccomplishedwith GMO bacteriaexpressing GFP (Green Fluorescent Protein) in presence of arsenic. • Our prototype willallow to quantify GFP i.e. arsenic concentration

  5. Whydevelop a new field kit? Pros & Cons

  6. Whatisour kit? • Communitybased / Understanding of the prototype • Reparable

  7. What about legal aspects? • The test kit would use genetically modified bacteria outside a qualified laboratory • The main issue is to have the GMO confinedfrom the environment. • Wethusrefer to the « Ordinance on Handling Organisms in Contained Systems» 814.912 • According to Art.7, our kit is of class 1 i.e. null or negligibleriskactivity.

  8. An example of approved GMO field kit: ARSOlux • Detection of arsenic via bioluminesence • Use of a freeze-dried GMO bioreporter • Samevial • Approved by the Germanauthorities • Tested in Bangladesh and other countries http://www.ufz.de/arsolux/index.php?en=20706 http://www.ufz.de/export/data/411/33860_111005_ARSOlux_Projektkennblatt_englisch.pdf Sigfried et al. Environ SciTechnol 2012 46(6) pp.3281-87

  9. What about confinement? • “contained system means a system that uses physical barriers or a combination of physical and chemical or biological barriers to limit or prevent contact between organisms and people or the environment “(Art.3,h.) • After use, Bacteria and antibiotic are destroyed in the lab by autoclaving or inactivated by hypochlorite solution.

  10. Details of the GMO arsenic reporter strain • K-12 E.Colibacteriastrain (similar to ARSOlux: • E. coli DH5α) • Can’t do conjugation due to missing F factor and deficientrecA (biologicalbarrier) • Plasmid Design • the plasmid carries kanamycin resistance, for easy selection

  11. RiskAnalysis • What happens if these GMO bacteria are released in the field? CouldKanamycinresistance and the arsenic reporting system bespread? -> This E. coli strain cannot transfer the plasmid by itself to other bacteria. Should cells lyse, the plasmid DNA can be liberated and theoretically be taken up by naturally transformable strains. • GFP non toxic and its fluorescence is not dangerous for the cell. • Other drugs exist to kill Kanamycin resistant bacteria. -> This resistance could be removed but the detection efficiency would decrease.

  12. Antibiogram

  13. Future Design & Possibilities • Turbiditymeasurement to normalize the GFP signal • Electronic system incorporatedinside the box • Built-in interface (buttons, screen…) • Use with a smartphone via an application • Adaptable to otherbioreporters for harmfulelementsdetection (Mercury etc…) • Field test

  14. Answer to possible questions

  15. Why As is not good? Where? • Interfere with Krebs cycle (inhibits pyruvate conversion to acetyl-coA) • As a slow poison, causes diseases • Skin diseases • Intestinal tract problems • Cancers • Maximum concentration advisedby WHO: 10μg/L • Letal dose: 1mg/kg/day • Problem in Bangladesh and someAsian countries

  16. Whatisourcurrentcost? • Around 50 CHF • Possibility to develop a cheaper prototype usinganother micro-controller, i.e a simplerArduino hardware.

  17. Why not detect bioluminescence?

  18. Differencebetweenarsenite and arsenate? • Arsenite has lessoxygenthanArsenate Arsenite AsIII (As2O3, AsO2-, AsO33-) Reporter more sensitive Arsenate AsV (AsO43-)

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