2007 Synthetic Biology Team Challenge March 19-23, 26 Instructors: Howard Salis, Jeff Tabor

Course Information Monday – Friday: 9AM-5PM Final Presentations: Monday 3/26 1:30-3PM GH 114 Course wiki:

Schedule Monday –Intro to Synthetic Biology –Lab: Registry of Standard Biological Parts Tuesday –Survey of useful parts –Journal Club –Lab: Engineer novel genetic logic Wednesday –Modeling gene networks in MATLAB (H. Salis) –Homework: Brainstorm synthetic system Thursday –Develop plan for system –Optimize system with model Friday –Specify system with appropriate parts from literature –Document parts and systems in the registry –Simulations of final systems Monday (1:30-3:00) –~15 min Final presentations. –5 Faculty judges decide winner –Winning group’s design is synthesized.

Outline - Monday 1.Brief History of Molecular Biology 2.Dawn of Synthetic Biology 1.Concepts driving early designs 2.Building genomes from scratch 3.Landmark efforts in system design 4.System talks 1.Liz Clarke 2.Dan Widmaier 3.Matt Eames 5.Abstracting/formalizing the design process 6.Lunch 7.Afternoon Lab. MIT’s registry of Standard Biological Parts.

Chronology of Molecular Biology Structure of DNA. Watson and Crick Concept of mRNA, Regulator/operator pairs, Operons. Jacob and Monod. -(Gene networks of any desired property can be assembled from combinations of simple regulatory elements) Discovery of codons and the genetic code Recombinant DNA technology (Cohen and Boyer, UCSF) DNA Sequencing Technology Invention of PCR. "Beginning with a single molecule of the genetic material DNA, the PCR can generate 100 billion similar molecules in an afternoon.“ –K. Mullis First Automated Sequencer (Applied Biosystems Prism 373) Sequence of E.coli genome published (Blattner, UW) Sequence of human genome published (HGP/Celera) CSI:Miami debuts on CBS

Synthetic Biology Nature 403, January 2000

Cells are composed of complex networks Adapted From: Lee et al., Science, 2004

Complex networks are composed of simpler modules

Modules can be reconfigured into synthetic networks Elowitz and Leibler. Nature, 2000

Simulating a synthetic gene network Continuous ModelDiscreet Model Elowitz and Leibler. Nature, 2000

Genetic Toggle Switch Gardner et al. Nature, 2000

Carlson, Pace & Proliferation of Biological Technologies, Biosec. & Bioterror. 1(3):1 (2003) c/o Drew Endy -DNA synthesis capacity has doubled each 1.5 years over the past 10 years -System design and fabrication can routinely be decoupled (Endy, Nature 2005)

Building genomes from scratch Assembly of functional poliovirus genome (~7.5kb; Cello et al., Science 2002). –Oligos designed computationally, ordered commercially –[C H N98245 O P7501 S2340] Assembly of a bacteriophage genome (~5kb; Smith et al., PNAS 2003). –2 weeks assembly time Assembly of the 1918 flu virus (~13kb). (Tumpey et al., Science 2005). Craig Venter’s Mycoplasma genitalium genome = 580kb

Rewriting genomes (Chan et al., Molecular Systems Biology, 2005)

wt refactored

Genetic pulse generator Basu et al., PNAS 2005 Sender E.coliReceiver E.coli

Genetic pattern formation circuit Basu et al., Nature 2005

2004 UT-Austin/UCSF Bugwarz Team Not pictured: Andy Ellington, Chris Voigt

High-resolution spatial control of gene expression Projector Agar plate

Engineering light-dependent gene expression in E. coli

Bacterial photography Wild-type EnvZ MaskBacterial lawn

“Light Cannon” Mercury vapor lamp Concave grating spectrometer Actuator Projected image 37 degree incubator Double Guass focusing lens 35 mm slide 632nm bandpass filter

Improved black and white photography Endyrichia coli Escherichia ellington

‘Biofilm’ capable of continuous expression response Levskaya et al., Nature, 2005

Continuous response allows capture of high information images

Bacterial edge detector Projector Agar plate

Genetic logic for edge detection Only occurs at edge of light/dark

Gates mismatched: LOW output from gate 1 interpreted as HIGH input at gate 2 Light repression is incomplete

Matching gates through RBS redesign

Contributed Talks 10:00-10:30: L. Clarke ‘A Bacterial Thermometer’ 10:30-11:00: D. Widmaier ‘Secreting Spider Silk in Salmonella’ 11:00-11:30: M. Eames ‘Remote Controlled Bacteria’

Genetic “Parts” for programming living cells Voigt, Curr. Opin Biotechnol., 2006

Genetic “devices” integrate signal inputs Voigt, Curr. Opin Biotechnol., 2006

Device outputs control “actuators” which determine cellular behaviors Voigt, Curr. Opin Biotechnol., 2006

Making Biology a reliable engineering discipline Standardization –Composability Characterization –‘off the shelf’ functionality Centralization –Well documented repositories Abstraction –Distribution of expertise/labor

Device characterization

stry/index.php/Part:BBa _F2620

Abstraction hierarchy for the engineering of biology Endy, Nature 2005

Lunch GH 114 sai