(665b) Transcriptional Control through Synthetic Genetic Regulation Devices in Clostridium sensu Stricto
AIChE Annual Meeting
Thursday, November 1, 2018 - 12:48pm to 1:06pm
In this work, we have developed two systems for transcriptional control in Clostridium sensu stricto. First, we have developed a regulatable CRISPRi gene repression for the fine-tuning of biosynthetic pathways using multiple Cas12a effector proteins, which are better suited for use in Clostridium due to the genusâ AT-rich genomes. We demonstrate tunable repression based on proximity to regulation elements, strand, and number of targeted sites through reporter genes, transcription levels, and metabolic alteration. We propose a set of heuristics for such control. In addition, we demonstrate a set of engineered product-responsive transcription factor regulated promoters (biosensors) for real-time single cell monitoring of butanol accumulation. We demonstrate tunability of gene expression though altering the number, location, and sequence of the transcription factor binding sites. Such TF biosensors can be used for isolation of hyperproducing mutants and rewiring metabolism during fermentation production.
 Joseph, R. C., Kim, N. M., and Sandoval, N. R. (2018) Recent Developments of the Synthetic Biology Toolkit for Clostridium, Front. Microbiol. 9.