Sponsors

ACKNOWLEDGEMENTS:-Sian Davies (Advisor)-William Rostain (Advisor)-Alfonso Rosales-Jaramillo (Supervisor)

-Support of the staff and technicians of Warwick Life Sciences in particular the mammalian group of David Evans -Leo Vong-Chelsey Tye

HASSAN (MATHS) IVA (ENGINEERING) CHRIS (PHYSICS) BECKY (BIOMED)

WAQ (BIOLOGY) CARRIE (BIOMED) DAN (MATHS) BEN (MATHS)

Wellcome trust Sanger Institute

CDC: cdc.gov/diabetes/pubs/factsheet11.html

Dipeptidyl Peptidase IV (DPP-IV) inhibitors

Efficacy

Cost

0

2

4

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12

Inhibtor 1 Inhibitor 2 Inhibitor 3

Pri

ce p

er t

able

t ($

)Price per Tablet ($)

Inhibitor 1

Dic

er

RISC

1. Slicing

2. Unwinds3. Complimentary annealing

Target mRNA (3’UTR of DPP-IV)

RISC4. Slicing

Interfering RNA Treatment

T7

5’ 3’

IN VITRO TRANSCRIPTION

RNA dependent RNA polymerase (RdRp)

3’ 5’

POSITIVE STRAND RNA

NEGATIVE STRAND RNA

5’ 3’POSITIVE STRAND RNA

REVERSE TRANSCRIPTION

3’ RNA PROMOTER

MS2 Repressor System

MS2 coat P2A RdRp

MS2 coat protein

RdRp

MS2 BOX

IRES

Aptazyme

• THEOPHYLLINE CHEMOSENSOR

• SELF-CLEAVING RIBOZYME

SWITCH

siRNA Engineering

DICERDICER

BIOLOGICAL DIODE

NEGATIVE SENSE RNA POSITIVE SENSE RNA

Promoter

Repressor

Selectivity Marker

Translational Apparatus

Kill Switch

Replication System

Active Element

The RNA Construct

Promoter

5’ RNA promoter

3’ RNA promoter

Split Neomycin Selectivity Used in selectivity for co-transformants when testing the siRNA and aptazyme in a separate module to RdRp.

Schmidt, C., Shis, D., Nguyen-Huu, T. and Bennett, M. (2012). Stable Maintenance of Multiple Plasmids in E. coli Using a Single Selective Marker.

AND LOGIC GATE

Kanamycin resistance in prokaryotes and genticin resistance in eukaryotes

Testing the Replication systemRNA DEPENDENT RNA POLYMERASE

BL21 E. coliCELLS

RBS +RdRp

RBS + reverse GFP + 3’ RNA Promoter

RdRp Results

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RdRP induced Negative ControlRdRP not induced

Negative Controlmutant RdRP induced

GFP

Flu

ore

sce

nce

RdRp Activity in E.coli BL21 at O.D. 0.2

FLU

OR

ESC

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E (A

.U.)

Internal Ribosome Entry Site Testing EMCV vs NKRF

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EMCV Huh EMCV Hela NKRF Huh NKRF Hela Negative CntrlHuh

Negative CntrlHeLa

Flu

ore

scen

ce (

A.U

.)

Comparison of efficacy of EMCV and NKRF IRES with controls

Aptazyme

IN VITRO FZ Huh no theo

IN VIVO

FZ Huh + theo FZ HeLa no theo FZ HeLa + theo

0mM4mM

20mM 100mM

-RNA degradation

+RNA production

-RNA production

-RNA production

MS2 Translation

+RNA

MS2

-RNA

siRNAdegradation

RISC Complex degradation

siRNA

RISC Binding

RISC Complex

DICER action

MS2 Binding

+RNA degradation

MS2 degradation MS2-RNA

Complex

MS2-RNA Complex Degradation

RNA Annealing

dsRNA

dsRNAdegradation

RdRp Translation

RdRp

RdRp Degradation

Further Modelling Developments

The time delay differential equations for our 3’RNA promoter experiment, solved numerically (in red) and stochastically (in blue, averaging 100 repeats) with the solver we created

Toolbox

Policy & Practices

SURVEY · BIOWEAPONS · PROJECT-SPECIFIC CHANGES · OUTREACH

Survey

Word of Mouth

The Internet

Other iGEM Teams

Viral Methods

#323

Intellectual Property IIf someone modifies an organism or cell such that it now has a new

function, do you believe that the person has right to claim ownership of this modified organism?

‘No one has the right to life’

‘Modifying cells/organisms in a particular way is its own unique work that should be as copyrightable as any

other invention’

YES48%NO

52%

Intellectual Property IIIf someone modifies human cells, or maybe even a human, to have a new/modified function, can that person claim ownership of these

human cells/human?

‘Ownership should only extend to the copyright over the modified segments of

genetic code. The organism itself, however, is under it's own ownership if it has sentience.’

‘Technically yes. Obviously you would pay to have a modification done on you. You're paying for both a service and a product. The "product" used to belong them, so technically they were the previous owners; but once someone buys

them, they are the new owners.’’

YES13%

NO87%

Associated DangersDo you believe Synthetic Biology is a dangerous tool, knowing that

potentially dangerous organisms are being dealt with?

‘Synthetic biology is a double edged sword. It has equally well advantages and disadvantages. As responsible scientists/policy makers, we need

to make sure that the technology is not being used to threaten life.’

YES30%

NO45%

OTHER25%

Confidence in SynBioWould you be happy taking medicine that is derived using bacteria

and viruses which have been selectively modified to serve as a cure for a particular illness?

YES94%

NO6%

BioweaponsMerriam-Webster definition: a harmful biological agent (as a

pathogenic microorganism or a neurotoxin) used as a weapon to cause death or disease usually on a large scale

Problem 1: using a malicious siRNAsequence

Problem 2: exploiting self-replication to promote a hazardous toxin

Outreach

Lucie’s testimony: It gave me a new appreciation for all areas of the research

and the intricacies of carrying it out.’

Outreach

The World of RNA