jamal elkhader & queenie chan 22 february 2012
DESCRIPTION
“Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology” Baojun Wang, Richard I Kitney, Nicolas Joly & Martin Buck. Jamal Elkhader & Queenie Chan 22 February 2012. Background. AND gate:. Background. NAND gate:. Background. Terms : - PowerPoint PPT PresentationTRANSCRIPT
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“Engineering modular and orthogonal genetic logic gates for
robust digital-like synthetic biology”
Baojun Wang, Richard I Kitney, Nicolas Joly & Martin BuckJamal Elkhader & Queenie Chan
22 February 2012
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Background
AND gate:
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Background
NAND gate:
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Background
Terms:o Digital system: on/offo Forward engineering: classifying/evaluating
components before choosing the parts to build the system
o Modular: can change inputs and outputs and logic gate will still work
o Orthogonal: no cross-talk
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Project Overview
Purpose: to engineer a digital-like system using modular and orthogonal parts via forward engineering
I. ClassificationII. Design AND gateIII.Test ModularityIV. repeat II and III for NAND gateV. Compatibility
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Classification: Forward Engineering
• 3 different promoters: Plac, PBAD, Plux
• 3 test conditions:1. promoters' responses, varying chassis (E. coli
MC4100, E. coli MC1061)
IPTG --> lacArabinose --> BADAHL --> lux
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Classification: Forward Engineering
• 3 different promoters: Plac, PBAD, Plux
• 3 test conditions:1. promoters' responses, varying chassis (E. coli
MC4100, E. coli MC1061)2. promoters + RBSs responses with varying RBS's
IPTG --> lacArabinose --> BADAHL --> lux
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Classification: Forward Engineering• 3 different promoters: Plac, PBAD, Plux
• 3 test conditions:1. promoters' responses, varying chassis (E. coli
MC4100, E. coli MC1061)2. promoters + RBSs responses with varying RBS's3. Promoters + RBS's responses with varying
temperatures
IPTG --> lacArabinose --> BADAHL --> lux
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Circuit Design: AND Gates
Gate 1
Gate 2
Testing for Modularity
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Gate 230ºC
37ºC
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NAND gates: Design & Modularity Test
Gate 3
Gate 4
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Gate 3 Gate 4Predicted
Results
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NAND gate FACS data
Both inputs added -->
off
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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• Compatibility of AND gate with chassis
• Top: Gate 1 (Plac + PBAD)
• Bottom: Gate 2 (Plux + PBAD)
Compatibility ✗✗ ✗ ✗
✗
✗
Classification Design AND gate
Test Modularity NAND gate Compatibilit
y
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Conclusions• AND and NAND gates are modular
• Sigmoidal response property allows system to be robust (minimal background noise)
• Forward engineering and engineering-by-parts works!
• Crosstalk can be avoided by picking promoters that are not native to chassis
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SignificanceModular:
• Can use other inputs and outputs and AND / NAND gate will still work
• Parts-based engineering approach
Control:
• New genetic logic devices can be engineered predictably to generate desired behaviours in response to specific extra- or intra- cellular signalling inputs
• Improves sensing specificity and accuracy of biological control
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Supplementary Slides Plac
PBAD
Plux