A CRISPR-Cas9 tool to explore the genetics of Bacillus subtilis phages.
Lett Appl Microbiol
; 71(6): 588-595, 2020 Dec.
Article
em En
| MEDLINE
| ID: mdl-32615024
ABSTRACT
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.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Bacillus subtilis
/
Fagos Bacilares
/
Sistemas CRISPR-Cas
Idioma:
En
Revista:
Lett Appl Microbiol
Ano de publicação:
2020
Tipo de documento:
Article