Protocol for genomic recombineering in Yersinia ruckeri using CRISPR Cas12a coupled with the λ Red system

Research output: Contribution to journalJournal articleResearchpeer-review


  • Fulltext

    Final published version, 4.1 MB, PDF document

Genomic manipulation of Yersinia ruckeri, a pathogen of salmonid fish species, is essential for understanding bacterial physiology and virulence. Here, we present a protocol for genomic recombineering in Y. ruckeri, a species reluctant to standard genomic engineering, using CRISPR Cas12a coupled with the λ Red system. We describe steps for identifying protospacer guides, preparing repair template plasmids, and electroporating Yersinia cells with Cpf1 and protospacer plasmids with homologous arms. We then detail procedures for genome editing and plasmid curing.

Original languageEnglish
Article number103014
JournalSTAR Protocols
Issue number2
Number of pages17
Publication statusPublished - 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors

    Research areas

  • Biotechnology and bioengineering, CRISPR, Genomics

ID: 390184224