Synthetic Biology: Biopharmaceuticals & Insulin-
generating Enteric Bacteria
Vi Nguyen
Biopharmaceuticals & MDR-TB/XDR-TB
What Are Biopharmaceuticals?
• Medical drugs created using biotechnology
• Include:– interferons– hormones– clotting factors– vaccines– antibodies
Tuberculosis
• caused by Mycobacterium tuberculosis
• 8.7 million contracted TB in 2011
• 630,000 cases of MDR-TB– resistant to most powerful tuberculosis drugs
Synthetic Gene Circuit for Drug Screening
Going Further
Bibliography"Expanding Nature’s Toolkit: How Synthetic Biology Is Changing the Face of
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World Health Organization. "Tuberculosis (TB)." WHO. United Nations, 2013. Web. 7 July 2013. <http://www.who.int/topics/tuberculosis/en/>.
Insulin-generating Enteric Bacteria (IGEBs)
Purpose
• Provide a more convenient means of insulin therapy for diabetics
• Modify native gut flora to produce insulin (E. coli)
• Bacteria that produce insulin at ideal times (during glucose intake)
Competing Technologies
• Insulin injections– Pros:
• Relatively inexpensive• Relatively simple to
administer
– Cons:• Requires daily injections• Blood glucose spikes• Insulin resistance may occur in repeated needle
stick areas
Competing Technologies
• Insulin pumps– Pros:
• More accurate doses• Fewer blood glucose spikes• More flexible lifestyle• Fewer needlesticks
– Cons:• Expensive• Bulky system constantly attached to body• Requires extensive training to use
Design
• Determining when to produce large amounts of insulin
• No glucose no insulin– Glyoxylate cycle in absence of glucose
• Modified quorum sensing– Produce large amounts of insulin at certain
times
SuccinateSignaling molecule
Repressor Insulinproduction
LuxS
lsr promoterLuxS TAT peptide export signal
INS
succinate
AI-2 signaling molecules
ABC transporter
ATP ADPATP
ADP
lsr transport cassette
LsrR
P
LsrRP
TAT export
prepoinsulin
insulin molecules
AI-2
insulin
ATP
ADP
Expected Results
• During times of carbohydrate intake insulin production by IGEBs
Glucose Insulin production
0 0
1 1
Glucose Insulin production
absent 150 units
present 2000 units
Advantages
• Fewer required treatments
• Completely internal system
• Self-adjusting system
• Very flexible lifestyle
Potential Problems
• Surviving gastrointestinal tract
• Adhering to villi in small intestine
• Ensuring adequate absorption of insulin
• Horizontal gene transfer?
Testing
• Insulin production in absence/presence of glucose in environment– Cells exposed to various cycles of glucose
absence and presence– Insulin production measured and tracked over
time
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Taqa, ME, JL Semmelhack, and BL Bassler. "The LuxS-dependent Autoinducer AI-2 Controls the Expression of an ABC Transporter That Functions in AI-2 Uptake in Salmonella Typhimurium." Molecular Microbiology 42.3 (2001): 777-93. PubMed.gov. Web. 9 July 2013. <http://www.ncbi.nlm.nih.gov/pubmed/11722742>.