Dr. Haoqian Zhang

Dr. Haoqian Zhang:

When we first came up with the idea of synthesizing the precursor of active ingredient of catnip, nepetalactol, we faced tremendous difficulties; we don’t know clear applications of nepetalactol and questioned ourselves about the value of it to the society; also, it is quite esoteric for a high school iGEM team to construct genetic circuits inside yeast, which none of our team members have any experience with. With these questions, we visited Dr. Haoqian Zhang, the CTO of Bluepha Co., Ltd.

Professor Christopher Voigt

Professor Christopher Voigt:

Having decided to use TALEsp in our project, we were not sure about whether should we apply it to yeast or not. Thus on July 7th, at the 4th Synthetic Biology Youth Scholar Forum, we asked professor Christopher Voigt about his opinion on integrating TALEsp into yeast and other technical questions related. Professor Voigt,, the leader of the team who created TALEsp, hinted that since TALEsp sequences are similar to each other, an application in yeast would most possibly trigger the reassemble system of yeast and dysfunction its ability. Also, in the aspect of E.coli TALEsp, he suggested integrating the circuits to the genome to prevent failure of TALEsp when a large number of bacteria exist (e.g fermenter). When coming into the technique of characterizing TALEsp promoters, we were uncertain of which method to use: GFP or RNA-seq. He clarified that these two techniques are not of much difference, only that RNA-seq is a little more accurate, but at the same time much more expensive. We thus decided a GFP characterization method.

GreatBay_China with Professor Voigt

Dr. Qiuli Lou

Dr. Qiuli Lou:

To work with yeast is a great challenge to our team. Many information of Yeast remains unclear, yet the most emergence one is that we don’t know exactly the condition to carry out colony PCR. To tackle this problem we sought help from Lou, a graduate student majoring on biotechnology at Fujian Agriculture and Forestry University. From this conversation, we figured out a way of diluting free colonies in 20mM NaOH for 10min 100˚C before the normal PCR process. Breaking through this technical problem our yeast circuits construction gets to move on.

Dr. Junbiao Dai

Dr. Junbiao Dai:

Our design on experiment involves knock out of 4 enzymes and knock in of 3 enzymes in yeast, and we primarily decide, in the knock out experiments, to use the Hi-crisper technique developed by Junibiao Dai and his team. (further details see our design page: ) However, after several attempts, we found the technique troublesome: they did not work as demonstrated by the paper. Also, during the expression stage, we found out that E.coli often dies when our pYES plasmid is transformed, yet yeast often will not carry this plasmid when transformed directly from Gibson product. We thus sought for a chance to interview Dr. Dai and inquired him the problem we faced. He suggested that if Hi-crisper will not work, then a traditional knock out process is recommended to be carried out. Also, for the dying E.coli plight, he hinted that the pYES plasmid with three coding sequences of the enzymes to be expressed might be toxic to E.coli as a whole. The circuits heavies the metabolic burden of E.coli tremendously and leading to their death. Finally, he debugged the yeast issue, saying that Gibson product has a low concentration, and presented a feasible solution of lengthening the homogenous arm to trigger the homogeneous recombination stem of yeast. We adapted his suggestion of knock out sequence one by one.

Dr. Chong Zhang

Dr. Chong Zhang:

Chong Zhang is the partner of Junbiao Dai; he introduced he and Dr. Dai’s paper: MiYA, an efficient machine-learning workflow in conjunction with the YeastFab assembly strategy for combinatorial optimization of heterologous metabolic pathways in Saccharomyces cerevisiae to us. This paper introduced a possible way to enhance the production of nepetalacol in yeast, and we grew very keen on adopting this method. However, due to the lack of database and the scanty of time, this method was obsoleted.

Dr. Akihiko Kondo

Dr. Akihiko Kondo:

We met Dr. Kondo at the 4th Synthetic Biology Youth Scholar Forum and introduced our project to him, with whom grew very interested. He suggested his point mutation plasmid to us. Due to the lack of time, we did not integrate this suggestion into our project; but we are thankful to his help and appreciate his kindness.

We participated the 4th Synthetic Biology Young Scholar Forum, where we met Dr. Akihiko Kondo. We gained a lot of knowledge and advice there and broadened our scope

Fenghua Li:

Fenghua Li:

We managed to contact a stray cay rescue figure in Beijing, named Fenghua Li. Having pursued this activity for 20 years, first on her own then with her rescue team, she hinted us that stray cats are of extreme alert and is often hard to capture. In order to save more stray cats and to eliminate the future population of them, the process of TNR (Trap, Neuter, and Release) is essential. This reminds us of a construction of a ‘Kitty wonderland’ (KW) to which cats are attracted when our nepetalactol is placed in, and which can aid the work of the rescue team. Besides that, acting as a household, if possible, to stray cat population. Associating with our project of nepetalacol, the proposal not only suggests a way of applying our project, but also brings benefit to the whole society by eliminating the population of stray cats. From then on, our first draft and prototype of this device are proposed, and the campaign of our hardware begins.

Rescue Team:

Rescue Team:

To improve our KW, we decided to construct a 3D model; but whether is this device really aiding the works of rescue teams or not, or if it's but a burden to them, remains unanswered. Thus a few weeks later we managed a thorough online interview with the leader of the rescue team of Ms. Fenghua Li, Dachui, and introduced for the first time our thoughts of this KW to the stakeholder. To our surprise, this design was much approved by Dachui and other team members, and consolidated our will of materializing the KW.

Li Yu

Li Yu:

When the first draft was schemed, we sought professional help from specialists in X.factory, Shenzhen’s first and leading maker space since 2011. By then we were extremely unconfident with our design, and kept on delaying the finale of our 3D model. When we talked through our situation with Li Yu, a designer in X.factory, she urged us to carry out our first prototype and put it in practice.