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Selected Case Studies QuickTime™ and a TIFF (LZW) decompressor are needed to see this pict ure. MIT 2006: Engineering bacteria to smell good C. breweri S. cerevisiae BSMT ATF1 wintergreen banana Slides borrowed from the 2006 MIT Team Engineering the scent of bacteria QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. What is the desired output? Components of engineered cell QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. What is needed to create the desired output? Components of engineered cell QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. What is needed to create the desired output? Creating devices with BioBricks Protein Regulatory Part/ Protein Coding Part Promoter Ribosome Binding Site Terminator Components of scent-producing devices QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. Components of scent-producing devices QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. Components of upstream devices QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. How do you regulate when the scents are expressed? QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. Goal: Sense the growth state of culture QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. Regulating the timing of expression osmY: active in stationary phase & under high osmotic pressure conditions osm Y osmY + inverter Regulating the timing of expression QuickTime™ and a TIFF (LZW) decompressor are needed to see this picture. 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 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 Ribosome Binding Site Protein Coding Part Regulatory Part/ Promoter Protein Terminator DNA to RNA to Protein DNA Ribosome Binding Site RNA Ribosome Binding Site Protein Protein synthesis from RNA Ribosome RNA Ribosome Binding Site Protein Locked structure prevents ribosome binding Ribosome RNA Ribosome Binding Site Protein RNA “Key” unlocks lock Ribosome RNA Ribosome Binding Site Protein Goal: Create network of cellcell communication 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