TY - GEN A1 - Otte, Kolja A1 - Kühne, Nora A1 - Furrer, Alexandra A1 - Lozada, Lina A1 - Lutz, Veronika A1 - Schilling, Tobias A1 - Hertel, Robert T1 - A CRISPR-Cas9 tool to explore the genetics of Bacillus subtilis phages T2 - Letters in Applied Microbiology N2 - Here we present pRH030, a new CRISPR-Cas9 tool for the genetic engineering of Bacillus phages and beyond. It is based on the Streptococcus pyogenes cas9 with its native constitutive promoter, tracrRNA, and a gRNA precursor. The constitutive expression of Cas9 was conducive to the inactivation of viral attackers and enhanced phage mutagenesis efficiency up to 100%. The gRNA precursor can be built-up to an artificial CRISPR array with up to 5 spacers (target sequences) assembled from ordinary oligonucleotides and directly cloned into pRH030. Required time and resources remain comparable to a single gRNA cloning. These properties make pRH030 an attractive new system for the modification of Bacillus phages and qualify it for research beyond genetic construction. Y1 - 2020 UR - https://opus4.kobv.de/opus4-UBICO/frontdoor/index/index/docId/25856 UR - https://sfamjournals.onlinelibrary.wiley.com/doi/abs/10.1111/lam.13349 SN - 1472-765X VL - 71 IS - 6 SP - 588 EP - 595 ER -