Ilea's collaborative project with Joe Bondy-Denomy's lab on genome engineering using CRISPR-Cas3 has been published in Nature Methods! A challenge for functional genomics in Pseudomonas syringae is to create strains that lack many or all effectors, which are critical for virulence of the pathogen. The Collmer lab created an effectorless strain of P. syringae by removing every effector gene through homologous recombination, however this approach is not easily scalable for multiple strains. We showed that CRISPR-Cas3 could make large deletions of clusters of effector genes. We demonstrate that the CRISPR-Cas3 mutants behave like traditionally constructed mutants from the Collmer lab, which validates this approach. This will be a powerful tool for gene characterization in multiple species of bacteria, including P. syringae. The article is available here: https://www.nature.com/articles/s41592-020-00980-w
Posted Sept 30, 2020, updated Oct 20, 2020.