Programmable cells: Interfacing natural and engineered gene networks
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Transcript of Programmable cells: Interfacing natural and engineered gene networks
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Programmable cells: Interfacing natural and engineered gene networks
Hideki Kobayashi, Mads Kærn, Michihiro Araki, Kristy Chung, Timothy S. Gardner, Charles R. Cantor, and James J. Collins
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• create novel cellular behaviors and characteristics by coupling engineered gene networks to the cell’s natural regulatory circuitry
• Four examples• Detects and retains memory of DNA damage• Forms biofilm in response to DNA damage• Detects and retains memory of quorum sensing
molecules• Density dependent protein synthesis
Scope
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Flow Cytometer (BD FcsCalibur)
Reporter: GFP
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lacI -> lacR -> Ptrc -> cI -> cI -> PL
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lacI -> lacR -> Ptrc -> cI -> cI -> PL
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MMC – 15 hUV – 1-10 s
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(With traA gene) (lacking the traA gene)
Replacegfp with traA
Biofilm was only observed if traA was expressed for > 4h
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LuxI-LuxR quorum-sensing systems
acylated-homoserine lactone (AHL)
luxICDABEluxR
LuxR LuxI
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Quorum sensing molecules
lacI -> lacR -> Ptrc -> cI -> cI -> PL
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Density-dependent gene activation
Toggle
lacI -> lacR -> Ptrc -> cI -> cI -> PL
GFP
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Discussions
• Programmable cells have been constructed by interfacing natural and engineered gene networks
• Programmable cells have been demonstrated using four different constructs with the toggle gene network as a building block
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Something to think about…
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DNA damage sensing
b)
Original design
Will the modified design work? If not, why not? If yes, how would it differ from (a)?
Modified design
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Density-dependent gene activation
Q: What if we replace the lacI gene with gfp and forget about the regulatory circuit
gfp
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