1. Physical insulation of pathways or functions

2. How to build complex synthetic circuits? Electronic hard wired: reusability of components. Biology is diffusive: crosstalk We need to find a way to insulate the different components circuits.

3. Pathway insulation methods Orthogonality (chemical insulation) Family of orthogonal parts Physical insulation Scaffolding Synthetic organelles Synthetic ecosystems

4. PROJECT 2: BUILDING A NEW ORGANELLE FOR SYNTHETIC BIOLOGY STEP 1: Create spatially distinct membrane compartment that has unique molecular identity wild-type cellcreate “synthesome”use “synthesome” STEP 2: Recruit proteins to carryout any synbio process of choice (e.g. drug or biofuel factory) A quick recap

5. An external organelle External organelles producing cell Targetting Components of a pathway to secretion External organelles producing cell

6. Chassi s (Feeder) External organelle An external organelle

7. Chassi s (Feeder) Attachment transmembrane proteins An external organelle

8. External organelle Chassi s (Feeder)

9. External organelle Chassi s (Feeder) communication

10. External organelle composition (1) Chassi s (Feeder) factory Receiver sender Chassis (Feeder) Function?

11. Advantage: part reusability Chassi s (Feeder) Chassis (Feeder) State output Example: multicolor Hasty oscillator output

12. External organelle composition (2) Chassi s (Feeder) pathway Starting product

13. Natural example: Nanoarchaeum equitans

14. Nanoarchaeum: ABC transporters Chassi s (Feeder) Communication: Within eukaryotes, ABC-transporters mainly transport molecules to the outside of the plasma membrane or into membrane-bound organelles such as the endoplasmic reticulum, mitochondria, etc.

15. Gap junctions Bacterial conjugation Natural examples