Chances and limitations when uncovering essential and non‐essential genes of <scp> Bacillus subtilis </scp> phages with <scp>CRISPR‐Cas9</scp>
In: Environmental Microbiology Reports, Jg. 13 (2021-08-31), S. 934-944
Online
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Zugriff:
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.
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Chances and limitations when uncovering essential and non‐essential genes of <scp> Bacillus subtilis </scp> phages with <scp>CRISPR‐Cas9</scp>
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Autor/in / Beteiligte Person: | Hertel, Robert ; Muhammad M Nawaz ; Basu, Syamantak ; Kohm, Katharina |
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Zeitschrift: | Environmental Microbiology Reports, Jg. 13 (2021-08-31), S. 934-944 |
Veröffentlichung: | Wiley, 2021 |
Medientyp: | unknown |
ISSN: | 1758-2229 (print) |
DOI: | 10.1111/1758-2229.13005 |
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