BACTOBLOOD

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BACTOBLOOD
ResearchersArthur Yu Austin Day David Tulga
Hannah Cole Kristin Doan Kristin Fuller
Nhu Nguyen Samantha Liang Vaibhavi Umesh
Vincent Parker Teaching AssistantsAmin
Hajimorad Farnaz Nowroozi Rickey
Bonds Advisors John Dueber Christopher
Anderson Adam Arkin Jay Keasling
UC BERKELEY

• Creating a Red Blood Cell Substitute

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Artificial Blood Substitutes

• The Need

• Supply shortage, especially in
• developing countries
• PFC limitations
• HBOC limitations

Benefits of Bactoblood

• Universally compatible
• Disease-free
• Inexpensive
• Ability to be stored for a prolonged period
• Rapid production in emergency situations

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Human Practice IP Considerations
What makes Bactoblood novel and non-obvious? the
functional integration of all the devices into a
single system
What is patentable the part or the application
of the part? the combination of parts that
provide a function (device)
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Human Practices IP Considerations
What makes Bactoblood novel and non-obvious? the
functional integration of all the devices into a
single system
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Human Practices IP Considerations
What makes Bactoblood novel and non-obvious? the
functional integration of all the devices into a
single system
Patentability of Bactoblood may depend on what
aspects of the invention are claimed in a patent
application how it is worded.
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The Chassis
Protect E.coli from Immune System
Protect Recipient from E. coli
Lipopolysaccharide (LPS)
K1O16 capsule
Pili and Flagella
tonB gene
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Expression of Human Hemoglobin
Dimeric HbA
Monomeric HbA
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System Components
Alpha Hemoglobin Stabilizing Protein
Heme
Antioxidants
Cytochome b5 / Cytochrome b5 Reductase
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Freeze Drying

• Bactoblood can be stockpiled and easily
  transported
• 2 desiccation devices which prevent cell damage

Trehalose
Hydroxyectoine

• Four genes from
• Streptomyces chrysomallus

• 2 genes from e. coli genome

Both help cells recover after freeze-drying
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Freeze Drying
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The Controller
Directs copy number and transcription of system
devices

• T7 Polymerase
• pir genes
• Iron-inducible promoter

• Biosynthetic Operons
• T7 Promoters
• pir dependent R6K Origin

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The Controller
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Controller Part Characterization
T7 RNA Polymerase
Iron Promoter, yfbE

• Only composite part with the weakest rbs and a
  GTG start
• codon showed iron-dependent GFP production

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Copy Number Device Assays
GFP Cytometry As copy number increases, so
does the amount of GFP
Low copy number
High copy number
Iron-dependent copy number
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Genetic Kill Switch

• Prevents chance of infection or unwanted
  proliferation
• When induced, cells degrade their own DNA

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Kill Switch Growth Assays
of colonies
Arabinose
Arabinose
- Arabinose
- Arabinose
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Phenotype of Dead Cells
With Arabinose
Without Arabinose
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A Comprehensive System
Oxygen Delivery Peroxide Damage Control Survival
in Bloodstream Inability to Replicate

• yes
• yes
• yes
• yes

• Universal Compatibility
• Ability to be Freeze-dried
• Self-replicating
• Disease Free

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Acknowledgements

• The Arkin and Keasling Labs
• Kate Spohr, Kevin Costa and Gwyneth Terry
• SynBERC
• The Camille and Henry Dreyfus Foundation

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Patent Timeline
How might synthetic biology be a driver for
inventing new modes of industrial practices and
partnerships other than the current open source
approach?
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Swarming Assay
Wild Type (with flagella)
Chassis (no flagella)
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Serum Survival Assay
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Oxygen Transport
Oxygen Delivery
1 AU
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Oxygen Transport
Oxygen Delivery
25 AU
100 AU
P50 Value
O2 Saturation
pO2 (Torr)
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Problems
O2
Superoxide
Methemoglobin
Doesnt Work

Free Radicals
O2
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2
Not Good
O2
O2
O2