CRISPR-Cas System-Mediated Genetic Modification in Bacillus spp.: Current Status and Future

Area of Science:

• Microbiology
• Biotechnology
• Synthetic Biology

Background:

• * *Bacillus* species are Gram-positive bacteria with significant industrial potential.
• * They efficiently convert low-cost substrates into high-value compounds for feed, pharmaceuticals, and food.
• * Genetic modification is key to enhancing their production capabilities.

Purpose of the Study:

• * To review CRISPR-Cas systems applied to *Bacillus* species.
• * To outline strategies for CRISPR-Cas-mediated genetic modification in these bacteria.
• * To discuss expression regulation for optimizing *Bacillus* as cell factories.

Main Methods:

• * Review of CRISPR-Cas systems and their application in *Bacillus* genetic engineering.
• * Analysis of genetic modification strategies, including homologous recombination and base editing.
• * Examination of expression regulation techniques for metabolic engineering.

Main Results:

• * CRISPR-Cas technology significantly enhances the production performance of *Bacillus* strains.
• * Homologous recombination is a primary strategy, with base editing emerging as a precise alternative.
• * Effective expression regulation is crucial for developing efficient *Bacillus* cell factories.

Conclusions:

• * CRISPR-Cas systems offer powerful tools for the genetic improvement of *Bacillus* species.
• * Strategic implementation of these systems, alongside expression control, is vital for industrial applications.
• * Addressing current technical challenges will enable the development of superior genetically modified *Bacillus* production strains.