This year, we selected Chlamydomonas reinhardtti as the chassis organism. We knock down the genes related to lipid production, namely LACS1, LACS2, PLA2, ACX2, PEPC1, DTH1, and CHT7 genes in its metabolic pathway by utilizing the CRISPR/CAS9 system, so as to achieve the purpose of efficient lipid production.

Develop C. reinhardtii CC-503 culture method: during algae cultivation and mutant selection, we found that pollution from unknown microbes is severe. We change the antibiotics into newly developed cephalosporins and Timentin instead of Ampicillin, which is restricted by many bacteria in our lab.For more detailed information, please check our Lab Work page in the experiment part.

Perform lipid metabolic-centered transcriptome analysis: We use 7 different pressure conditions (Deprivation of nitrogen, sulfur, iron and potassium in diverse combinations) to culture C.reinhardtii and then extract RNA for transcriptome analysis to discover differential expressing genes linked to lipid production.For more detailed information, please check our Lab Work page in the experiment part.

We designed 12 Basic Parts and 5 Composite Parts, as well as optimizing the parts from previous iGEMers. Significant also, we compare fluorescent proteins mCherry and StayGold for selection and consequently construct 2 plasmids by using 2 different fluorescent protein genes and 2 diverse promoters for gRNAs, followed by comparing their function during genetic engineering of C.reinhardtii. Please check more details on the Parts page.

In our project, C. reinhardtii was gene edited byCRISPR/Cas9 system to achieve optimal lipid accumulation. Consequently，searching asuitable sgRNA is of pivotal significance. To gain a deeper understanding of the physical mechanisms behind gene editing, we develop a thermodynamics-based model to predict the potential off-target activity of the selected guide RNA. And the model we use overcomes the above binary limitations of the previous physics-based binary off-target predicting model and considers off-targets effects with both high possibility and low possibility.

We designed a card game called Algal Lab . To make the synthetic biology related knowledge more intriguing and accessible, we created an educational game called algal lab, which intimates the whole process of utilizing algae to produce bio-fuels in the lab. Also, we write a brochure for the game in great detail, including related synthetic biology knowledge and play regulations. This game has a total of 124 cards , including 24 algae cards, 30 culture medium cards, 30 function cards, 16 lipid cards, 24 property cards and 12 dice. For details, please click here to view our card game in detail.

To retain precious feelings and ideas in the process of participating in iGEM as well as benefiting the subsequent iGEMers, we wrote a brochure with our partner HainanU_China collectively. Please click here to view our brochure.