(188d) Tunable Crispri-Based Transcriptional Control in Clostridium Pasteurianum Using dCas12a
AIChE Annual Meeting
Monday, October 29, 2018 - 3:30pm to 5:00pm
Here we have developed a system for CRISPRi based repression in Clostridium using catalytically dead Cas12a (dCas12a) proteins from Moraxella bovoculi and Francisella novicida. We use a bioinformatics approach to show that these effectors are ideal for use in Clostridium based on the genusâ AT-rich genomes. We demonstrate repression in Clostridium pasteurianum, a solventogenic species able to produce a variety of chemicals including butanol from a wide range of feedstocks. We also show tunability of this system based on inducibility of dCas12a, proximity of target region to regulation elements, and strand targeted by quantifying transcription levels and solvent production. This system represents a very valuable addition to the metabolic engineering toolkit for Clostridium and would allow further progress in the optimization of various species of this genus for industrial applications.