A Dynamic, Modular, High Binding Affinity Scaffold for the Colocalization of Proteins and Control of Intracellular Metabolic Flux

TechnologyDescription: Researchers at the University of Delaware in Newark, DE have designed a newinnovative process that can add more dynamic control to synthetic metabolons.The scaffold consists of two components: proteins of the CRISPR/Cas Type 1Family, and short scaffolding RNA sequences. For the co-localization scaffold,Csy4 (P. Aeruginosa) and Cse3 (E. Coli) are the two proteins that areused. The hybridization regions of two scaffold RNAs can bind to each other,which shows that the fused proteins are successfully co-localized. Todisassemble the scaffold, a third RNA strand invades one of the scaffold’sgRNAs, and through toehold mediated strand displacement it pushes out thesecond scaffold gRNA and the scaffold dissociates. This disassembly step givesthe synthetic metabolons increased control.Patent Status: This method is patent pending. Further information on licensingopportunities is available on request.Application: This is a prospective new innovative process that can potentially add anextra level of dynamic control to synthetic metabolons, making them moresimilar to native metabolons. This could allow for greater product titer andimproved culture survivability. Problems Addressed: Generally, when producing small bio-chemicals such as pharmaceuticals, enzymesare introduced into a metabolic pathway of interest in the model organism. Thishas been successful for small chain hydrocarbon molecules which are in theaforementioned metabolic pathway. The problem with this process is that it isenormously tedious and also yields extremely low optimization of the turnoverand expression rate of the introduced enzyme. This low optimization is requiredin order to minimize the stress the new pathway inflicts on the model organism.To counter this problem, synthetic metabolons are used to increase the fluxthrough organized proteins via a process called substrate channeling. Theproblem with these synthetic metabolons is that they are unable to assemble anddisassemble on cue. Although product yield is optimized with these syntheticmetabolons, once they have been assembled, they offer minimal control over themetabolic flux distribution. After the synthetic metabolon assembles, it canonly redirect metabolic fluxes once. There is no such negative feedback loop asthere are in native metabolons. This invention can potentially give this samelevel of control that native metabolons possess to synthetic metabolons aswell.

United States

Patent Pending

USA