1. Selected Case Studies

2. MIT 2006: Engineering bacteria to smell good BSMT wintergreen C. breweriS. cerevisiae ATF1 banana Slides borrowed from the 2006 MIT Team

3. Engineering the scent of bacteria What is the desired output?

4. Components of engineered cell What is needed to create the desired output?

5. Components of engineered cell What is needed to create the desired output?

6. Creating devices with BioBricks Regulatory Part/ Promoter Protein Coding Part TerminatorRibosome Binding Site Protein

7. Components of scent-producing devices

9. Components of upstream devices

10. How do you regulate when the scents are expressed?

11. Goal: Sense the growth state of culture

12. Regulating the timing of expression osmY: active in stationary phase & under high osmotic pressure conditions osm Y osmY + inverter

13. Regulating the timing of expression

14. Design principles What is the desired output? What is needed to create this output? What do you want to sense? Use of constitutive promoters, inducible promoters, and inverters

15. Berkeley 2006 Bacterial Networks Need: To transfer DNA messages from one bacterial cell to another Means: Bacterial Conjugation Need: To specifically control who can read the DNA message Means: Riboregulation Some slides borrowed from the 2006 Berkeley Team

16. Regulatory Part/ Promoter Terminator Protein Protein Coding Part Ribosome Binding Site

17. Ribosome Binding Site Protein DNA RNA Ribosome Binding Site DNA to RNA to Protein

18. Protein synthesis from RNA Protein RNA Ribosome Binding Site Ribosome

19. Locked structure prevents ribosome binding Protein RNA Ribosome Binding Site Ribosome

20. RNA “Key” unlocks lock Protein RNA Ribosome Binding Site Ribosome

21. Goal: Create network of cell- cell communication

22. RNA as a regulatory mechanism Can activate some “riboswitches” with small molecules Advantage: fast response - transcript is already made Alternative method: block translation by adding antisense RNA to RBD