A CRISPR-Cas-associated transposon system for genome editing in Burkholderia cepacia complex species

• DOI: 10.1128/aem.00699-24
• Publication Date: 2024-06-13T13:01:39Z
• Authors: Zhong Ling Yap; A. S. M. Zisanur ...; Andrew M. Hogan; David B. Levin; Silvia T. Cardona
• Keywords: Transposase; Burkholderia cepacia complex; Burkholderia cenocepacia; Insertional mutagenesis; Forward genetics; Transposon mutagenesis

ABSTRACT Genome editing in non-model bacteria is important to understand gene-to-function links that may differ from those of model microorganisms. Although species the Burkholderia cepacia complex (Bcc) have great biotechnological capacities, limited genetic tools available and mitigate their pathogenic potential hamper utilization industrial applications. To broaden for Bcc species, we developed RhaCAST, a targeted DNA insertion platform based on CRISPR-associated transposase driven by rhamnose-inducible promoter. We demonstrated utility system insertional mutagenesis strains B. cenocepacia K56-2 multivorans ATCC17616. showed RhaCAST can be used loss- gain-of-function Importantly, selection marker could excised reused allow iterative manipulation. The faster, easier, more adaptable than previous disrupt pathogenicity elements insert relevant modules, enabling IMPORTANCE Species but are also opportunistic pathogens. Genetic manipulation necessary links. However, manipulate Bcc, hindering our understanding traits tool increase species. this study loss gain function significance work expanding currently Bcc.