valencia upv igem 2018 team · 2019. 12. 2. · 1 2 3 barriers to access synbio the access is...

1 2 3

Barriers to access synbio

The access is hampered by the required equipment, knowledge

and economic costs.

Human Practices

Hardware

Printeria combines multiple technologies to automatically produce genetic circuits.

1 Entry zone

BioInk (Bioparts)The BioInk is positioned in a carousel-based system, so a needle produces the fall of the assembly reagents.

Components• Carousel system• Peristaltic pump

• DNA basic pieces

• BsaI endonuclease, T4 DNA ligase

• Enzyme Buffer + BSA

• Destination vector

2 Reaction zone

Experimental and simulation results

ComponentsFor drop movement• Printed Circuit Board (PCB) • Surface: PTFE foil and

hydrophobic gelFor changing temperatures• Power resistor (37ºC zone)• Peltier effect (16ºC zone)

3 Output system

Here, the optimal conditions for growth and measuring Optical Density (OD) and Fluorescence (F) are set.

Components• Shaker• 650nm Laser• 480nm led• Light sensor

Extracellular AHL

Intercellular AHL (A) Lux R (R)

Lux R·AHLR.A

(Lux R·AHL) dimer(R.A) 2

gGFP

GFP

mGFP

TRANSCRIPTIONTRANSLATION +

LuxI promoter Plux

mR

An intuitive and user-adapted interface allows the user to design and print genetic circuits by selecting the basic DNA pieces.

Before printing a genetic circuit, it can be simulated with our Simulation Tool, or it can be directly printed.

These pieces are part of our Golden Gate standardized Part Collection, and they are fully characterized to provide the user all the information about the designed genetic circuit.

An extensive repository of pre-designed recipes for genetic circuits can be printed too.

Software

Printeria uses the Level 1 Golden Gate assembly method, so the multipartite directional assembly of a composite part (transcriptional unit) is carried out by a high robust one-step reaction in a drop.

The destination vector is introduced in its linearized form to avoid the screening of non-recombinant transformed plasmids.

With our ODE modeland the experimental data we have estimated model parameters and characterized all the parts in our Printeria part collection.

Applying different voltages to the pads in the PCB surface, local hydrophobicity changes. This makes the droplets move between the hot and cold zones.

For each Printeria recipe we take information from our database and combine it to get a ODE model of the new part. The user can predict the behavior of the design before printing.

Level 1 Golden Gate

Droplets movement

525

500

475

450

425

4000 50 100 150 200 250 300

Lux R

1800

1600

1400

1200

1000

8000 50 100 150 200 250 300

Protein of Interest

15k

12.5k

10k

7.5k

5k

2.5k

00 50 100 150 200 250 300

Number of cells

Simulation tool

Thermal image of the PCB showing the temperatures of the different zones

Calibration system

Parts characterization

Printeria Experiments

Printeria’s goal is to democratize SynBio and make it accesible to everyone: BioArtist, Research Labs, Teachers, and Students.

Integrated HumanFeedback from students (with the Kano Model) and renowned scientists and artists was taken into account in order to improve our device’s functionalities.

Social EngagementWe understand BioArt as the point where art and science meet, with art being the cornerstone of dissemination.

Educational approach to SynBio.

Yturralde and his Impossible Figure

María Peñil andMehmet

Carol from Printeria’s Team with Francis Mojica

GFPmut3b / GFPmut3b + LVA tag

Changing RBS and promoter strength

TU

BBa_K2656105

BBa_K2656106

BBa_K2656107

BBa_K2656117

BBa_K2656108

sfGFP

Changing RBS strength

TU

BBa_K2656101

BBa_K2656100

BBa_K2656102

BBa_K2656103

BBa_K2656104

mRFP1 / mRFP1 + LVA tag

Changing RBS strength

TU

BBa_K2656109

BBa_K2656118

BBa_K2656119

BBa_K2656110

Blue chromoprotein

Changing RBS strength

TU

BBa_K2656113

BBa_K2656120

YFP / YFP + LVA tag

TU

BBa_K2656112

BBa_K2656111

The calibration curve shows experimental data measured by Printeria’s sensors vs. the flourescein molecules number per cell (MEFL

GFP/cell) from our

Interlab Study.

Calibrated Optical Density curve made by Printeria’s OD sensor.

MEF

L GFP

/cel

l

Prot

ein

(MEF

L GFP

/cel

l)

Valencia UPV iGEM 2018 Team