Possibility of using CRISPR-Cas9 for inactivating genes controlling cell differentiation pathways in Bacillus subtilis

Bacillus subtilis possesses multiple differentiation pathways, all of which are activated when the cell population reaches stationary phase in culture. These differentiation states include sporulation, competence development, and cannibalism. Except for competence development which facilitates the insertion of DNA plasmid into the host cell, sporulation and cannibalism are not favourable from a biotechnology standpoint. Specifically, sporulation negates the ability of the cells to perform biocatalytic reactions for either biocatalysis or metabolic engineering applications. On the other hand, cannibalism reduces the cell population drastically; thereby, presenting a hurdle to growth coupled biosynthesis of products. Hence, a B. subtilis strain not able to enter into any of these differentiation pathways would be a better producer of value-added products in metabolic engineering and biocatalytic applications. One possibility of generating such a strain would be to use CRISPR-Cas9 to inactivate the genes involved in controlling entry to a variety of cell differentiation pathways.