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. 2023:2555:205-212.
doi: 10.1007/978-1-0716-2795-2_15.

CRISPR-Cas9 Shaped Viral Metagenomes Associated with Bacillus subtilis

Katharina Kohm  1 , Veronika Theresa Lutz  2 , Ines Friedrich  3 , Robert Hertel  4   5
Affiliations

CRISPR-Cas9 Shaped Viral Metagenomes Associated with Bacillus subtilis

Katharina Kohm et al. Methods Mol Biol. 2023.

Abstract

Phages are viruses of bacteria and have been known for over a century. They do not have a metabolism or protein synthesis machinery and rely on host cells for replication. The model organism Bacillus subtilis has served as a host strain for decades and enabled the isolation of many unique viral strains. However, many viral species representatives remained orphans as no, or only a few, related phages were ever re-isolated.The presented protocol describes how a CRISPR-Cas9 system with an artificial CRISPR-array can be set up and used to discriminate abundant and well-known B. subtilis phage from a host-based metagenome enrichment. The obtained viral suspension can be used for metagenome sequencing and isolating new viral strains.

Keywords: Bacillus subtilis; Bacteriophage; CRISPR-Cas9; CutSPR; Metagenome; Phage; gRNA.

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