Pseudomonas putida Genome Editing Services

Pseudomonas putida KT2440 and closely related P. putida backgrounds are the usual starting point. Other Pseudomonas hosts, engineered derivatives or client-supplied isolates are reviewed during scoping, because plasmid maintenance, selection markers and growth behaviour can differ between backgrounds and affect which editing route is practical.

Not usually. Knockout projects are typically run with donor-independent editing routes, which keeps construct assembly and screening simpler. When a project needs a defined sequence change, a donor template or an equivalent repair strategy may be required, and that decision is confirmed with you during the design stage.

Candidate clones are screened by locus-specific PCR and then confirmed by Sanger sequencing across the target region. Depending on the agreed scope, additional checks such as targeted amplicon analysis of nominated sites, plasmid-clearance testing or wider sequencing can be incorporated, and the confirmation plan is written into the project scope before work starts.

Plasmid curing and marker handling are addressed as part of the project scope rather than assumed. Where curing is required for downstream work, the approach is agreed at the design stage and the plasmid status of each clone delivered is recorded in the handoff documentation.

In silico prediction of candidate off-target sites is part of routine design work and the predicted sites are listed for your review. Experimental off-target testing, such as targeted amplicon checks of nominated sites, is added when a project scope calls for it; broader genome-level checks are agreed case by case.

Scheduling is agreed once scoping is complete. Single-target knockouts, multiplex panels and iterative design rounds are planned as separate milestones, and the confirmed schedule is issued with the project plan together with the decision points where you review screening and genotyping results.