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Chances and limitations when uncovering essential and non-essential genes of Bacillus subtilis phages with CRISPR-Cas9.
- Source :
-
Environmental microbiology reports [Environ Microbiol Rep] 2021 Dec; Vol. 13 (6), pp. 934-944. Date of Electronic Publication: 2021 Aug 31. - Publication Year :
- 2021
-
Abstract
- Virulent bacterial viruses, also known as phages or bacteriophages, are considered as a potential option to fight antibiotic-resistant bacteria. However, their biology is still poorly understood, and only a fraction of phage genes is assigned with a function. To enable the first classification, we explored new options to test phage genes for their requirement on viral replication. As a model, we used the smallest known Bacillus subtilis phage Goe1, and the Cas9-based mutagenesis vector pRH030 as a genetic tool. All phage genes were specifically disrupted, and individual survival rates and mutant genotypes were investigated. Surviving phages relied on the genome integrity through host intrinsic non-homologues end joining system or a natural alteration of the Cas9 target sequence. Quantification of phage survivors and verifying the underlying genetic situation enables the classification of genes in essential or non-essential sets for viral replication. We also observed structural genes to hold more natural mutations than genes of the genome replication machinery.<br /> (© 2021 The Authors. Environmental Microbiology Reports published by Society for Applied Microbiology and John Wiley & Sons Ltd.)
Details
- Language :
- English
- ISSN :
- 1758-2229
- Volume :
- 13
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Environmental microbiology reports
- Publication Type :
- Academic Journal
- Accession number :
- 34465000
- Full Text :
- https://doi.org/10.1111/1758-2229.13005