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Engineered bidirectional communication mediates a consensus in a microbial consortium. Presented by: Iman Mirrezaei. Outline. Introduction Problem definition Methods Experiments. Why am I still Sick . 170 Million American have Chronic disease What is Chronic bacterial disease?

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engineered bidirectional communication mediates a consensus in a microbial consortium

Engineered bidirectional communication mediates a consensus in a microbial consortium

Presented by: ImanMirrezaei

outline
Outline
  • Introduction
  • Problem definition
  • Methods
  • Experiments
why am i still sick
Why am I still Sick
  • 170 Million American have Chronic disease
  • What is Chronic bacterial disease?
    • Infection of bones
    • Make people suffer and make people die
  • The real number are staggering
    • 550,000 deaths annually
    • 30x the number of Aids patients
    • Equal to the number of cancer death
    • $94 Billion in medical costs
biofilm
Biofilm
  • A biofilm is any group of microorganisms
  • The ability to manipulate these films would enable controlled studies of microbial ecosystem dynamics

Staphylococcus aureusbiofilm

biofilm formation
BioFilm formation
  • Reversible Adsorption of Bacteria (sec.)
  • Irreversible attachment of bacteria (sec.-min.)
  • Growth and division of bacteria (hrs.-days)
  • Exopolymer production and BioFilm formation (hrs.-days)
  • Attachment of other organism to BioFilm( days- months)
biofilm relationship
BioFilm relationship
  • The BioFilm could maintain different relationship with the hosts:
    • Mutualism: both benefit
    • Commensalism: one benefit one unaffected
    • Parasitism: one benefit, one harmed
    • Competition: neither benefit
    • Neutralism: both unaffected
biofilms properties
BioFilms properties
  • Difficult to be Removed and Killed
  • Function as a single organism
microbial consortia form
Microbial consortia form
  • Microbial consortia form generate a function that none is capable of alone.
    • global food chain to human weight gain
  • A microbial consortium is two or more microbial groups living symbiotically
problem definition
Problem definition
  • Two colocalized populations of Escherichia coli converse by exchanging lactone signals.
  • This consensus function can be considered a logical AND gate
  • The ability to manipulate and control these films

Escherichia Coli

engineered circuits
Engineered circuits
  • Engineered circuits have been used to control the behavior of single cells and cell populations
  • Engineered communication
    • Broadcasting
    • Small-molecule signals
experiments requirements
Experiments Requirements
  • Engineered circuits have been used to control the behavior of single cells and cell populations in both time and space
  • Escherichia coli Bacterium
the mcc signaling network
The MCC “Signaling Network”
  • Components of the LasI/LasR and RhlI/RhlR quorum sensing systems found in Pseudomonas aeruginosa, that forms a biofilm in the lungs of cystic fibrosis patients

cystic fibrosis patients

modeling to minimize crosstalk
Modeling to Minimize Crosstalk
  • Minimize densities for consensus activation, maximize densities for isolation activation
  • Positive feedback loop
  • Need C4HSL for LasI and 3OC12HSL; need 3OC12HSL for RhlI and C4HSL (minimize crosstalk signal)
liquid culture
Liquid Culture
  • GFP( target gene) in both
  • Separate chambers with passage of small molecules
  • Responses over 100-fold greater with communication

Circuit A (with B)

Circuit B (with A)

Circuit B

(alone)

Circuit A (alone)

solid culture
Solid Culture
  • Cell types embedded separately in solid medium
  • Media placed in contact
  • Fluorescence decreases with distance from interface
  • Circuit A slower growth
biofilms consortium
Biofilms (consortium)
  • Circuit A – green (yellow); Circuit B – red (cyan)
  • Grow together and display MCC function (≥6dys)
  • No significant fluorescence separately
biofilm communication in nature
Biofilm Communication in Nature
  • Quorum sensing to coordinate biofilm formation
  • Biofilm disruption
  • Fundamental Applications
    • Integrate cells sensing different stimuli (temp, pH, ...)
    • More communication partners
    • Communication other than quorum
applied applications
Applied Applications
  • Mixed-culture batch reactors require quorum of each
  • Disrupt pathogenic BioFilms
  • Enzyme-prodrug pair; inactive toxin fragments
  • Materials synthesis
  • Surveillance for environmental changes
    • Epidemiology, material degradation
references
References:
  • 1. Hooper LV, Midtvedt T, Gordon JI (2002) Annu Rev Nutr 22:283–307.
  • 2. Kato S, Haruta S, Cui ZJ, Ishii M, Igarashi Y (2005) Appl Environ Microbiol

71:7099–7106.

  • 3. Kleessen B, Blaut M (2005) Br J Nutr 93(Suppl 1):S35–S40.
  • 4. Macfarlane S, Woodmansey EJ, Macfarlane GT (2005) Appl Environ Microbiol

71:7483–7492.

  • 5. Mishra S, Jyot J, Kuhad RC, Lal B (2001) CurrMicrobiol 43:328–335.
  • 6. Becskei A, Serrano L (2000) Nature 405:590–593.
  • 7. Becskei A, Seraphin B, Serrano L (2001) EMBO J 20:2528–2535.
  • 8. Kobayashi H, Kaern M, Araki M, Chung K, Gardner TS, Cantor CR, Collins JJ (2004) Proc Natl Acad Sci USA 101:8414–8419.
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