This year, we handed in 16 high-quality, well documented bio-brick parts, including all those we used in our project this year, and several brilliantly designed others forming a inspired by our iGEM project within these two years.
Truncated Trim21 (Part: BBa_K3396007)
It is the core of the newly registered Predator Pro system. Trim 21 comes from the family of TRIM. We truncated the Trim21 protein to maintain its RING domain, B box domain and coiled coli domain, while the antibody binding PRYSPRY domain was removed. This truncate protein maintained the E3 ubiquitin ligase activity, and provided an open interface for other protein dimerization pairs to be added.
TRIM21-Replaceable-1-P2A-Replaceable-2-Replaceable-3 (Part: BBa_K3396017)
Our favorite composite part BBa_K3396017 is a plasmid platform on which different targeting domains and protein dimerization pairs can be easily installed into the Predator Pro system. The subpart BBa_K3396007 functions as the E3 ligase to degrade target protein. The protein region (Part: BBa_K3396013) and (Part: BBa_K3396014) can be replaced into different protein dimerization pairs in order to achieve signal responsiveness. The (Part: BBa_K3396015) part can be replaced by other targeting domains (ScFv. Nanobody, ligand, etc.) to mediate targeted degradation. The parts BBa_K3396013- BBa_K3396015 were designed with flanking BsaI, BsmBI and AlwI sites for the scarless replacement of these parts.
As a member of the collection Predator system, special designs were taken for to optimize the applicability and adaptivity of such parts. Specifically, a novel designed substitution system, through which, two proteins could be fused with their corresponding fragment at the same time using Golden-Gate Assembly, was introduced to dramatically simplify the cloning process.
Figure 1.Schematic representation of the part.