During the progress of our project, we realised that “Inheritance” is important for every teams, especially for a new teams and the new iGEMers. Therefore, we sorted out some files and the data of parts, hoped that could inspire or give assistance to the future teams and iGEMers. Because of that we used CRISPR/Cas technology and cell free system, we make a CRISPR application guidance with teams including Tianjin-China, BUCT-China and Tongji-Software while we also compile Cell Free system handbook with ZJUT-CHINA. Furthermore, we uploaded several models and related data to GitHub. Moreover, we also completed the experimental characterization of the previous parts (BBa_C0062) and added the data to the corresponding part registry page.
Cell Free system handbook
Cell free system, or in vitro transcription and translation system, is a great choice for biological chassis chosen, as most of the construction process will met some problems on transformation or transfection. Meanwhile, it contains the advantages including high efficiency and relatively relaxed reaction conditions. Therefore, we make this Cell Free system handbook for the teams who are using or preparing to use it in their project. In this handbook, we collect the background of different cell free system and the principle of them. We also provided the detailed protocol of these kits. In the end, we compared these kits depending on our experience and experimental data.
CRISPR application guidance
As we all know, CRISPR/Cas is a popular efficient technology to edit the gene in vitro or in vivo. But because it is not mature enough, and it is new for many iGEM teams and iGEMers, it is frequently to meet various problem during the application of CRISPR/Cas system. In this year, several teams involve us made a CRISPR application guidance. The content includes the basic principle introduction of CRISPR/Cas system and numerous problems we met with the possible solutions provided by other teams or experts.
Model on GitHub
In this year, we construct several modelling to predict and simulate various biological progress including the infection of phage and the phage’s proteins expression (more detail on Modeling page). We uploaded these models and related data to GitHub, which could be used by other teams in the future to simplify their complex work.
Part: pLuxR promoter (BBa_R0062) measurement
In this year, we characterized pLuxR promoter by this part. As the Figure 1 shows, the composite part consists of the promoter pLuxR (K4054017), the RBS (K4054017)，the coding sequence of eGFP (E0040) and the terminator K4054018. This part was expressed in Cell Free systemm (in vitro transcription and translation system)， which allowed us to measure the GFP expression level under different LuxR concentration. More detail please see the part BBa_K4054100 (http://parts.igem.org/Part:BBa_K4054100) page.