INTRODUCTION

To make sure our project was fully ethical and implemented in a fully ethical way, we decided to consult with people who work in science and ethics.

We paid a visit to the DG research and Innovation in the EU in Brussels, Belgium where we met with a member on the ethical board, Dr. Joana Namorado. She walked us through the current EU laws set in place about ethical science and what to consider when designing our project to make sure it was safe. Finally she helped us use the Ethics self-assessment guide for the Horizon 2020 Programme set forth by the EU, which we used as a framework to create an ethical and safe design of our project.

Horizon 2020 ethical assessment

What is horizon 2020?
Horizon 2020 is the 8th Framework Programme for Research and Technological Development within the EU, a funding programme that defines the direction of current research within the EU area. It puts the focus on sustainable innovation.

What is the Horizon 2020 ethical assessment?
A large focus is put on the ethics of any project funded by the EU and thus, guidelines were specified on how to construct as assess the ethical aspects of a research project. The horizon 2020 ethical assessment is composed of 11 main sections, which apply in different levels to the various fields of current research:

Human embryos and fetuses
Human Beings
Human cells and tissues
Personal data
Animals
Non- EU countries
Environment, Safety and Health
Dual Use
Exclusive focus on Civil applications
Potential Misuse of research results
Other ethical issues

As we did not work with animal or human samples or subjects, sections 1 ,2 ,3 and 5 were not applicable to our project. Furthermore, all of our research was conducted in France, so section 6 was not applicable either.

Summary

We focused on the parts that pertained to our project:

Personal data

We made sure our data collection from questionnaires, interviews, phone calls and video recordings were taken with full consent and were kept on a secure domain. This data will not be transferred and does not contain any personal data (sensitive or otherwise).

Environment, Health and Safety

All of our team members were trained to work safely in the lab and taught about how to handle hazardous material, minimise transfer of genetic material and contamination and understand safety procedures.
We designed our prototype to limit contamination.
We designed our bacteria to have a safety kill-switch, based on a cell lysis part from bacteriophage.
We considered the precautionary principle in relation to our project, and found that there was no adequate safety risk that should prevent us from undertaking the project.

Dual Use

We identified all of the potential dual uses of our materials and control tools and propose that proper vigilance be taken to ensure that our tools are not used for any dual use purposes. However, we also postulate that the technology is not currently the most convenient (cheapest,easiest )for all the dual use applications we identified, therefore we believe that our technologies are not of primary concern for the issues we identified.

Exclusive focus on Civil Applications

Our project focusses solely on Civil Applications, and have no military affiliations or funding.

Potential misuse of Research results

Although we did not design our project to be harmful in any way to the general public, we were made aware that the wide scope of applications means that our technology can be applied in a variety of fields, through our human practices where we talked to both community biologists and academic scientists.

Other Ethics issues

None.

Full Assessment

Here is the list of the 8 different sections presented in the Horizon 2020 Programme set forth by the EU.

1) Human embryo and foetuses
2) Human beings
3) Human cells and tissues
4) Personal data
5) Animals

6) Non-EU countries
7) Environment, health & safety
8) Dual Use

Section 1, 2, 3, 5 and 6 were not applicable to our project as we are not using or implementing our design in any human or animal subjects or samples and all of our research was conducted in Paris, France.

Section 4 - Personal data

To conduct market research for our project, we collected data from different sources both about fundamental researchers using our 3D tools as well as community biologists and fablab users about our bioprinter.

We based ourselves on the Data Protection Directive, a european directive adopted in 1995, where personal data is defined as "any information relating to an identified or identifiable natural person ("data subject"); an identifiable person is one who can be identified, directly or indirectly, in particular by reference to an identification number or to one or more factors specific to his physical, physiological, mental, economic, cultural or social identity;" (art. 2 a).

This states that all data shall:

be obtained and processed fairly and lawfully and shall not be processed unless certain conditions are met.
be obtained for a specified and lawful purpose and shall not be processed in any manner incompatible with that purpose;
be adequate, relevant and not excessive for those purposes;
be accurate and kept up to date;
not be kept for longer than is necessary for that purpose;
be processed in accordance with the data subject's rights;
be kept safe from unauthorised access, accidental loss or destruction;
not be transferred to a country outside the European Economic Area, unless that country has adequate levels of protection for personal data.

Therefore, we made sure that none of our data was transferred.
All our online market research is fully anonymised and is not linkable to the person and therefore is not personal data.
All video, skype, face to face interviews were done with full informed consent, either through written documents or through video recording of consent and were notified of how their input would be processed.
The data we collected does not include: sensitive personal data (classed as: health, sexual lifestyle, ethnicity, political opinion, religious or philosophical conviction), genetic information, tracking or observation. This information was not deemed necessary for our project and thus we did not pursue any of the topics that would require special processing.
Furthermore none of this data was further transferred and thus no additional legal requirements need to be met.

Section 7 - Environment, health & safety

Environment

Because our research outcomes (including optogenetic tools for fundamental research, biomaterials for synthetic biology and materials research and bioprinter device) are all aimed at staying on contained laboratory settings, they do not pose any imminent threat to the environment and have minimal risk of being exposed.

DESIGN OF PROTOTYPE
The prototype for the 3D printer is fully contained, with glass doors allowing for the control of contamination as well as for the incubation of the bacteria at a correct temperature. The bacteria is contained in either a glass or plastic receptacle within the machine, which thus limit contamination within the machine. The interior of the machine is to be fully de-constructable and can be cleaned with ethanol. The design also includes a HEPA filter which would remove 99.97% of particles that have a size of 0.3 µm. A handbook of cleaning care would also be provided which would specify optimal cleaning protocols to limit contamination. Finally, This prototype was designed to be used only in labs and community spaces with level 1 criteria of biosafety, thus limiting hazardous contact with civilians.

BACTERIAL DESIGN
Furthermore, we designed a cell lysis part in our cells to ensure that our genetically engineered cells could be quickly lysed and contained through chemical induction.

Health and Safety

Our lab work and materials are all classed under level 1 biosafety criteria, and all team members underwent safety training for working in a laboratory with a Level 1 Biosafety criteria. This training was conducted by the Biosafety officer for INSERM U1001 and followed the practices described by the WHO laboratory biosafety manual.
This included learning:

Using protective clothing and materials
Decontamination and disposal methods
Chemical handling and storing
Use of Machines and Equipment
Emergency procedures
Hygiene standards and appropriate attire in the lab

For Section 7, we considered the Precautionary Principle, detailed in Article 191 of the Treaty on the Functioning of the European Union, where three preliminary conditions must be met: identification of potentially adverse effects; evaluation of the scientific data available; the extent of scientific uncertainty.

Potentially adverse effects

We identified potential risks associated with our project and then evaluated them.

Risk one: Hazardous material and products

Calcium carbonate: According to the cdc - ‘Health effects of exposure to the substance have been investigated, but none has been found. Calcium carbonate exists in nature as mineral aragonite and calcite (as in limestone, chalk and marble).’
Poly-silicate: These materials have not found to be hazardous.
Polyhydroxyalkanoates: these materials have found to be of no real hazard.

Risk two: Contamination
Risk three: dangerous GMO organisms

Evaluation of Scientific data available

The project was based on a preliminary extensive literature research, where we looked at the previous uses of the biomaterial production and applications, optogenetic parts, RNA scaffolding and are referenced throughout our project. We also kept close contact with research groups working in the same or similar research areas, which informed us that there were no previous risk/ incidents to environment and/or health.

Extent of scientific uncertainty

All of the biomaterials used are naturally occurring and have also been previously synthesised in labs (1.,2.,3.) as well as in iGEM projects. Although the CARP genes that were implemented in the Calcium Carbonate production have not been previously used in synthetic biology and in e.coli, the material and genes have been fully characterised.

The optogenetic tools used in the transmembrane circuitry, including the FixJ light receptor and Cph8 have been used in many previous works and are fully characterised. The Dronpa photoswitchable protein has previously characterised for use with enzymes and have also been characterised individually and thus we have reasonable scientific certainty of the function when combined with a transcription factor. The mApple photoswitchable protein is a protein that has been characterised individually and has high homology to the protein caging protein Dronpa. Therefore we can say with reasonable scientific certainty that mApple engineering will not create any dangerous adverse effects. The hybrid promoters created were all previously modelled in silico and were based on a previous paper that follows the same methodology.

Section 8 - Dual use

Dual use is is defined as “designed or suitable for both civilian and military purposes”.
Calcium carbonate- used in some formulations of smokeless powders used for loading firearms. However this is needed in a dust form and is only around 1% of the overall ingredients.
Polysilicate- used as a armor in ship hulls.

IN THE LAB

Safety training and Lab practice
All members of the team underwent safety training for working in a laboratory with a Level 1 Biosafety criteria. This training was conducted by the Biosafety officer for INSERM U1001 and followed the practices described by the WHO laboratory biosafety manual. This included learning:

Using protective clothing and materials
Decontamination and disposal methods
Chemical handling
Use of Machines and Equipment
Emergency procedures
Hygiene standards and appropriate attire in the lab

IN OUR DESIGN

Circuit Design
We designed a cell lysis part that would be implemented in the cells and could be induced chemically. This ensures that our printer design would be safe to use in both a research laboratory setting and in community biology labs. The part comes from a Bacteriophage, the

Prototype Design

Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France

bettencourt.igem2017@gmail.com

Team:Paris Bettencourt/Safety