UoM iGEM | Project Cutiful

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Manchester iGEM 2019 has been brainstorming productive ideas since Christmas 2018, meeting with, and benefitting from, the work of many people in order to develop our synbio hair-dye: Cutiful. The result of over ten months’ work, Cutiful is a holistic hair care product able to colour, repair, scent, and bind to hair without opening the outer layer (cuticle), thus preserving the structure and health of the hair.

Our transformed E. coli cells have the ability to:

• Secrete green (sfGFP), blue (amilCP), and red (mRFP1) chromophores with, and without, hydrophobic tags (experiments performed by the Colour team, led by Elisa Barrow, with members: Camilo Albornoz and Jonathan Fung)

• Bind to hair shaft, (experiments performed by the Colour Team)

• Secrete limonene from Acetyl CoA via the GPPS-LimS pathway, (experiments performed by the Smell team, led by Sara ElSafadi, with members: Ewan Egan, Marva Chan and Owen Baugh)

• Secrete vanillin from tyrosine via the enzymatic Sam8-Sam5-COMT-FSC-ECH pathway, (experiments performed by Smell Team)

• Secrete decapeptide G with, and without, hydrophobic tag for infiltration into damaged cortex. (experiments performed by the Decapeptide team, led by Thomas Harrison, with members: Mujtaba Ansari – main actor of wet labs – and Sophie Guillemot).

Cutiful has also achieved the following:

• Several theoretical models: smell diffusion, kill switch, allergenicity, ‘plaster’ for allergen-tests, light absorption... which have guided our choices of chromophores, odorants and human practices, (modelling performed by Thomas Harrison, Mujtaba Ansari, and Sophie Guillemot)

• Several outreach activities to engage and educate the public about GMOs, iGEM, and hair dyes, (whole team involvement)

• Attendance at Symposiums and Meetups to gather feedback and advice on experimental wet-lab as well as human practices, (whole team)

• Design of multiple interviews and discussions to gather sociologically meaningful data to guide our project development, (Mujtaba Ansari, with help from Camilo Albornoz, Ewan Egan, and Owen Baugh)

• Handling of funds and ordering of lab materials, (Jonathan Fung, with help from Camilo Albornoz)

• Obtention of sponsorships, (lead by Ewan Egan, with contribution from Camilo Albornoz)

• Market studies, (Jonathan Fung, with contribution from Owen Baugh and Vinordas Petkus)

• And Wiki creation (functional design: Thomas Harrison; website designer and Wiki lead: Jonathan Fung; arts and aesthetics: Marva Chan, Sophie Guillemot, with help from Elisa Barrow in designing the Cutiful logo; Contents: Whole Team).

Manchester iGEM 2019: From left to right: Sophie Guillemot, Mujtaba Ansari, Jonathan Fung, Ewan Egan, Camilo Albornoz, Sara ElSafadi, Elisa Barrow, Thomas Harrison

Our supervisors, Professor Eriko Takano and Professor Rainer Breitling, affiliated with the Manchester Institute of Biotechnology, are to be thanked for giving us the opportunity to compete, setting us up with an office, lab access, safety and technical training (in the form of our Induction Week), and initial funding. They have challenged us to keep being better throughout the developmental process, and have demanded nothing less but the best we could offer. Profs. Takano and Breitling have helped tremendously with narrowing down our ideas for Cutiful, eliminating the unfeasible, and have struck the right balance in letting us shape the project freely. They have shown enthusiasm, support and the right amount of admonishing throughout the project, keeping Cutiful on-track without taking its realisation from the team’s hand, checking over results and approving fund distribution, helping us to secure outreach abilities and giving us access to helpful contacts. For such intelligent supervision and endless patience, a very big ‘thank you’!

We also owe a heartfelt ‘thank you’ to our instructors, Dr Gajendar Komati Reddy and Dr Daniel Schindler, for their help throughout the project. Gajendar trained us when we first entered the lab, during Induction week (June 10th 2019 – June 15th, contents available on our Wet Lab page, Induction Week entry), and both have since looked over our results, helped us interpret data and checked over lab protocols. As the main deliverers of our synbio knowledge (sole training arranged was Induction Week, though help with understanding the science has always been forthcoming), they have also helped us select feasible experiments and ideas from the multitude we had. We must also thank them for their good humour and relativity, helping us keep our feet on the ground and keep calm throughout the whirlwind the iGEM competition has been. (Their master class in friendly teasing has also been appreciated, and put to good use!)

Katherine Baker, PhD student, iGEM instructor for Manchester 2018, and member of the Takano Group, has been unfailing in her help, both in the lab and with presentation-related works. She has doubly helped, by giving our instructors challenging advice and support, passing on her own iGEM experiences as a student, an instructor, and a judge.

The Takano group (Oksana Bilyk, Harshwardhan Poddar, Katy and Gajendar – to name only a few) has laughed at us, watched us and helped us as we stumbled through wet-lab and experimental design. We thank them for their tips, advice and general good humour in keeping an eye on the ‘iGEM newbies’, as well as their patience in being our (sometimes unwilling) audience for presentation rehearsals.

We thank our primary sponsors, affiliated with the University of Manchester: the Manchester Institute of Biotechnology (MIB), the School of Biological Sciences (from the Faculty of Biology, Medicine and Health), and the Department of Chemistry (from the Faculty of Science and Engineering).

Bear donated by our BMG Labtech sponsors.

And our academic sponsor:

• Professor Colin Jefcoate, University of Wisconsin-Madison, who organised a donation of fund from the Jefcoate Foundation.

For their less frequent, but ever-helpful advice, we also thank the following:

Decapeptide advice:

• Dr Artur Cavaco-Paulo, from the University of Minho, Braga (Portugal), author of several papers referenced in our research, who approached us at the Newcastle Synbio Symposium and offered both experimental and theoretical advice.

Vanillin advice:

• PhD student Guadalupe Alvarez Gonzalez, from the MIB, whose previous work on vanillin enabled her to give us helpful advice on cloning enzymes from the vanillin pathway in E. coli.

• Dr Cunyu Yan, whose help with the GC-MS has made detection of the vanillin we produced possible

Modelling Support:

• From the MIB, Aliah Hawari, who helped Thomas with modelling bacterial growth, and Francesco Del Carratore, who checked over our models, before passing them on to Professor Breitling for final approval.

• Also from the MIB, Dr Adam Amara, whose advice on Genome-wide modelling helped Thomas shape his work.

• Chad Greene (University of Texas at Austin), David Lee, Kenneth Hobson, and Fordi, whose code were used and expanded upon by Thomas and Sophie in devising our models.

• The I-TASSER software, from Yang-Zhang Labs, which was used in modelling peptides docking to protein, and University of Manchester student Graeme Krantz, for his help with PDB files, enabled Mujtaba to look into allergenicity more closely and devise a theoretical ‘allergen-testing plaster’ for use by the public.

Lab Support:

• iGEM Sheffield 2019, whose design of a microplate reader enabled them to run some characterisations on our behalf, and contributed towards collaboration.

• Dr Stefan Hoffman who donated to iGEM some bacterial colonies to test for hair adhesion.

Technical training:

• Dr Pablo Carbonell, based in the MIB, who provided the Smell’s Team with the required software for codon optimisation of the enzymic pathway turning tyrosine into vanillin. We also appreciated his presence during presentation rehearsals, and the questions he posed us.

• Sandra Taylor, who graciously lent to our team a microplate software for as long as we needed it.

• Our sponsor BMG Labtech, who commissioned Oliver Carney and Umear Sheikh to train team members in the use of the microplate readers they kindly lent us.

Presentation Coaching:

• Obtained from Emilia Wojcik, whose patience and constructive criticism bolstered our spirits and complemented the Takano Group’s – notably Katy’s – help.

Human Practices:

Many people have helped us in developing Cutiful into a responsible, thoughtful product. From giving us a sociologist’s point of view, to helping us attend outreach activities, as well as developing our project through uncomfortable questions and sometimes brutally honest advice, we must thank the following for their help and support:

• Dr Nick Weise, in charge of outreach at the MIB, who offered us a stand at the Science Community Festival.

• Dr Robert Meckin, whose constant help has shaped our Human Practices’ outlook on Synbiophobiaphobia, and who has ensured our surveys, interviews and questionnaires were sociologically meaningful.

• The University of Manchester’s Museum of Natural Science and Ancient History, where the Science Community Festival was held and who welcomed us and our project.

• Dr Rosalind LeFeuvre, whose help in getting our poster printed for the Synbiochem Symposium in March enabled us to attend and received a plethora of useful advice.

• Dr Tanya Aspinall, responsible for Health and Safety at the MIB, for her help in wading through the legalese describing regulations on experiments performed on hair obtained from a living, consenting individual. She also delivered the compulsory Safety course team members attended, allowing us to work in the laboratories of the MIB.

• Liam Byrne, from the Institute of Trichologists, who asked us many pointed technical questions and patiently answered all of ours. His sensible remarks are at the core of many adjustments Cutiful’s human practices underwent.

• Seamus McCrory, a hairdresser who agreed to speak with us and gave us remarkable insight in the day to day handling of dyes by professionals, enabling us to obtain a third point of view from possible users of our product.

• the IP lawyers from Venner Shipley LLD, Paul Misselbrook and Matthew Handley, for their time spent in educating us on intellectual property, giving us the scaffolding needed to view Cutiful from a commercial perspective, and consider the possible future of Cutiful, beyond iGEM.

• Edinburgh OG iGEM 2019, whose project on dye degradation in the textile industry has overlapped with our own human practices and have agreed to collaborate with us on a survey.

• Newcastle iGEM 2019, who organised the National iGEM UK Meetup and enabled us to not only form friendships with the other teams, but also gain confidence in presentation, gather our peer’s feedback on our product and, of course, attend the symposium where we exchanged with Dr Cavaco-Paulo.

• Our five anonymous interviewees, who generously gave up their time to answer our questions.

IP seminar, at Venner Shipley LLD: Clockwise around the table, beginning bottom left: Sara ElSafadi, Jonathan Fung, Sophie Guillemot, Paul Misselbrook, Matthew Handley, Mujtaba Ansari, Thomas Harrison, Ewan Egan, Camilo Albornoz (Elisa Barrow behind the camera).

Finally, for their help in making us a successful iGEM team, we thank:

• Sarah Jones, from the MIB, in helping us book flights, buses and hotels.

• Manchester iGEM 2018, for giving us tips on how to handle finances, not to put off wiki development and general moral support. Their suggestions on the handling of innocuous day to day tasks has also been greatly appreciated.

• Manchester iGEM teams 2013, ’14, ’15, ’16, and ’17, who left all of their work on a very precious hard drive, enabling us to learn from their experiences, and whose helpful members are still, sometimes, dotted around the MIB.

Team:Manchester/Attributions

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