This part is the new composite fusion protein that holds together the different sub-units of Runella zeae Gasdermin (bGSDM), a pore forming bGSDM protein, allowing for a specific use. The E. coli can already produce the bGSDM endogenously, but our project part makes it possible for the cell to produce this specific bGSDM/Csx30 fusion protein complex. This fusion protein contains a motif that is targeted by the caspase-like protein (TPR-CHAT/Csx29) associated with the Type III-E RNA-targeting effector complex (gRAMP/Cas7-11) known as Craspase. In previous experiments, Csx29 of the TPR-CHAT/Csx29 complex targeted the Csx30 linker of a human Gasdermin fusion protein to cause pyroptosis in the presence of an oncogene.
Others may use this part in tangent with a Craspase complex to target a variety of genes with therapeutic purpose. Modifications will need to be made to adjust the gRNA of the gRAMP/CAS7-11 complex to complement the DNA sequence of the gene being targeted. Further modifications of this part will need to be made to ensure that it is usable in other organisms (e.g., the Gasdermin itself may need to be adjusted to ensure that it can be produced in the host organism - being mindful of size and transcription sites, ensuring linker inserted fits in a region that will not significantly debilitate the protein once cleaved).
Subparts: BBa_K4608005 (Proteus, McGill 2023) - we modified this subpart in order to facilitate efficient folding and translation in E. coli and to prevent misfolding. Our modifications include deletion of certain ribosome binding sites and utilized online modelling programs to determine the best place to insert our linker.
https://parts.igem.org/Part:BBa_K5202001