A Programmable CRISPR/Cas9 Toolkit Improves Lycopene Production in Bacillus subtilis
Yang Liu, Haijiao Cheng, Haoni Li, Yingzhe Zhang, Meng Wang
Abstract
We present a toolkit that allows for rapid cloning procedures and one-step subcloning to move from plasmid-based expression to stable chromosome integration and expression in a production strain in less than a week. The utility of the customized tool was demonstrated by integrating the MEP (2C-methyl-d-erythritol-4-phosphate) pathway, part of the pentose phosphate pathway (PPP), and the hetero-lycopene biosynthesis genes by stable expression in the genome. The tool could be useful to engineer B. subtilis strains through diverse recombination events and ultimately improve its potential and scope of industrial application as biological chassis.
Topics & Concepts
CRISPRBacillus subtilisCas9Terminator (solar)BiologyMetabolic engineeringGuide RNAGenome editingGeneTranscription activator-like effector nucleaseGreen fluorescent proteinLycopenePromoterComputational biologySynthetic biologyHeterologousLocus (genetics)Genome engineeringGeneticsGene expressionBiochemistryBacteriaAstronomyCarotenoidPhysicsIonosphereCRISPR and Genetic EngineeringRNA and protein synthesis mechanismsMicrobial Metabolic Engineering and Bioproduction