Protocol for genomic recombineering in Yersinia ruckeri using CRISPR Cas12a coupled with the lambda Red system
(2024) In STAR Protocols 5(2).- Abstract
- 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.
- Abstract (Swedish)
- 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 l 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.
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/a81e9dd2-8831-445c-9f17-f54fa7e22727
- author
- Ejaz, Rooshanie N.
; Taylor, Nicholas M.I.
and Steiner-Rebrova, Eva M.
LU
- publishing date
- 2024
- type
- Contribution to journal
- publication status
- published
- subject
- in
- STAR Protocols
- volume
- 5
- issue
- 2
- article number
- 103014
- publisher
- Cell Press
- external identifiers
-
- pmid:38615317
- scopus:85190088394
- ISSN
- 2666-1667
- DOI
- 10.1016/j.xpro.2024.103014
- language
- English
- LU publication?
- no
- id
- a81e9dd2-8831-445c-9f17-f54fa7e22727
- date added to LUP
- 2024-06-19 13:09:49
- date last changed
- 2024-06-20 08:25:07
@article{a81e9dd2-8831-445c-9f17-f54fa7e22727, abstract = {{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.}}, author = {{Ejaz, Rooshanie N. and Taylor, Nicholas M.I. and Steiner-Rebrova, Eva M.}}, issn = {{2666-1667}}, language = {{eng}}, number = {{2}}, publisher = {{Cell Press}}, series = {{STAR Protocols}}, title = {{Protocol for genomic recombineering in Yersinia ruckeri using CRISPR Cas12a coupled with the lambda Red system}}, url = {{http://dx.doi.org/10.1016/j.xpro.2024.103014}}, doi = {{10.1016/j.xpro.2024.103014}}, volume = {{5}}, year = {{2024}}, }