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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

The CRISPR toolbox for the gram-positive model bacterium Bacillus subtilis

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Zocca, Vitoria Fernanda Bertolazzi [1] ; Correa, Graciely Gomes [1] ; Lins, Milca Rachel da Costa Ribeiro [1] ; de Jesus, Victor Nunes [1] ; Tavares, Leonardo Ferro [1] ; Amorim, Laura Araujo da Silva [1] ; Kundlatsch, Guilherme Engelberto [1] ; Pedrolli, Danielle Biscaro [1]
Total Authors: 8
[1] Univ Estadual Paulista UNESP, Sch Pharmaceut Sci, Dept Bioproc Engn & Biotechnol, Araraquara, SP - Brazil
Total Affiliations: 1
Document type: Review article
Web of Science Citations: 0

CRISPR has revolutionized the way we engineer genomes. Its simplicity and modularity have enabled the development of a great number of tools to edit genomes and to control gene expression. This powerful technology was first adapted to Bacillus subtilis in 2016 and has been intensely upgraded since then. Many tools have been successfully developed to build a CRISPR toolbox for this Gram-positive model and important industrial chassis. The toolbox includes tools, such as double-strand and single-strand cutting CRISPR for point mutation, gene insertion, and gene deletion up to 38 kb. Moreover, catalytic dead Cas proteins have been used for base editing, as well as for the control of gene expression (CRISPRi and CRISPRa). Many of these tools have been used for multiplex CRISPR with the most successful one targeting up to six loci simultaneously for point mutation. However, tools for efficient multiplex CRISPR for other functionalities are still missing in the toolbox. CRISPR engineering has already resulted in efficient protein and metabolite-producing strains, demonstrating its great potential. In this review, we cover all the important additions made to the B. subtilis CRISPR toolbox since 2016, and strain developments fomented by the technology. (AU)

FAPESP's process: 18/24733-0 - Development of a photo-activated kill switch device for biocontainment of genetically modified microorganisms
Grantee:Victor Nunes de Jesus
Support type: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 14/17564-7 - Reprogramming the purine metabolism of Bacillus subtilis employing sRNA-technology
Grantee:Danielle Biscaro Pedrolli
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 20/16058-1 - Optimization of a glucose-responsive genetic circuit
Grantee:Leonardo Ferro Tavares
Support type: Scholarships in Brazil - Scientific Initiation