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Physical insulation of pathways or functions 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. Pathway insulation methods Orthogonality (chemical insulation) Family of orthogonal parts Physical insulation Scaffolding Synthetic organelles Synthetic ecosystems PROJECT 2: BUILDING A NEW ORGANELLE FOR SYNTHETIC BIOLOGY A quick recap wild-type cell create “synthesome” use “synthesome” STEP 1: Create spatially distinct membrane compartment that has unique molecular identity STEP 2: Recruit proteins to carryout any synbio process of choice (e.g. drug or biofuel factory) An external organelle External organelles producing cell Targetting Components of a pathway to secretion External organelles producing cell An external organelle External organelle Chassi s (Feeder) An external organelle Attachment transmembrane proteins Chassi s (Feeder) External organelle Chassi s (Feeder) External organelle Chassi s (Feeder) communication External organelle composition (1) Function? Receiver factory Chassis (Feeder) Chassi s (Feeder) sender Advantage: part reusability Example: multicolor Hasty oscillator State output Chassis (Feeder) Chassi s (Feeder) output External organelle composition (2) pathway product Starting product Chassi s (Feeder) Natural example: Nanoarchaeum equitans QuickTime™ and a BMP decompressor are needed to see this picture. 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, Natural examples QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. Gap junctions QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. Bacterial conjugation