- We registered our team in the 2016 iGEM competition, worked very hard all summer and will attend the Jamboree at Boston!
- We have completed the judging form
- We received the deliverables through the post
- We have created a page on our team wiki, where we have attributed each aspect of the project, clearly distinguishing our work from that of advisors and mentors as well as acknowledging sponsors.
- We have contributed a collection of 18 parts, among which is BBa_K2148010 (mRaspberry fluorescent protein), which has been documented on the registry, establishing the phytobrick standard in iGEM (first chloroplast part in the competition!). We hope this will kickstart the use of plastid synthetic biology among future iGEM teams!
- We have validated our parts, submitted to the registry by sequencing and/or restriction digests.
- Experimental validation of aadA (BBa_K2148001 officially in registry) as seen through bacterial transformation.
- Possible to make L1 constructs for the experimental validation of parts through biolistic transformation of CW15 C. reinhardtii strain.
- We have also confirmed the correct insertion of all our 18 parts, either by sequencing of restriction digest. Check out wiki for more information on validation of hardware and software projects!
- Collaborated with Imperial iGEM team by providing them with Chlamydomonas growth data, for use with their computer model A.L.I.C.E.
- We established a close relationship with LiU iGEM team, also working on C. reinhardtii, by sharing weekly reports and skype meetings to follow up on each other’s work and advice on protocols.
- Attended and presented at the EU-funded Bio-Design Night science hosted in Paris along with the iGEM Paris-Bettencourt team.
- Strongly engaged in the topic of public perception and involvement. Presented our project at the EU-funded Bio-Design Night science conference as well as the interdisciplinary Co-Lab workshop.
- Engaged in discussion about the ethic of synthetic biology at the Co-Lab workshop, involving participants from 10+ countries. As a part of the workshop, conducted a survey of public opinion on the topic.
- Organized an outreach event on synthetic biology at a team member’s previous school.
- Several members were also actively involved in the inauguration of a Cambridge BioMakespace as well as the Cambridge University Synthetic Biology Society.
- Have discussed our experimental design with over 15 experts in the field, mainly on the issue of biological containment, our homoplasmy strategy and the impact of the project.
- Connected with the Cambridge Centre for Global Equality on the potential of plastid engineering for the use in developing countries and have integrated their advice in the design of our homoplasmy strategy.
- For our open-source hardware (biolistics and growth facility), have consulted 7 community science groups, from 5 countries, to make our design as much user-oriented as possible.
- Have submitted a version of an existing Cas9 part, which we codon-optimised for C. reinhardtii chloroplasts, among other codon-optimised parts.
- Cas9 was also optimised for the Phytobrick standard by removing illegal restriction sites with silent point mutations
- Have modified Cas9 overhangs and removed the stop codon for the assembly of a FP-fusion in a one-step reaction with a fluorescent protein (VFP) from our library.
- Have also submitted a version of the GFP gene, which we also codon-optimised.
- Have demonstrated our project under real-world conditions, i.e. in the chloroplasts of C. reinhardtii, as opposed to E. coli, where some of our parts could also be verified (due to the similarity of the two expression systems). To do so, have assembled two full gene units using our modular library, which were transfected into CW15 C. reinhardtii and conferred resistance to two different antibiotics.
- Proved that our library of C. reinhardtii chloroplast L0 parts (first-ever in iGEM!) can be used to assemble higher-order constructs, using the recently adopted Phytobricks standard. The assembly of the resulting L1s was confirmed with a restriction digest.