Microbiosci is a leading biotechnology company offering the best Salmonella genome editing services. With years of experience and expertise in microbial genome editing, our talented scientists will work closely with you to provide any help in Salmonella genome editing services.
Salmonella is the foodborne pathogen with the biggest public health and economic impact globally. It's one of the major causes of diarrhea all around the world, responsible for an estimated one million deaths and 100 million human infections annually. Genomic information decoded from a multitude of different Salmonella strains and new dramatic insights into pathogenic processes emphasize the fact that Salmonella research is currently at a very exciting juncture.
Salmonella Genome Editing Based on Homologous Recombination
Red/ET Recombination permits the engineering of DNA in Salmonella using homologous recombination mediated by phage protein pairs, either RecE/RecT or Reda/Redb. The central step in Red/ET recombination is the crossover step between a targeting construct containing homology arms and the target which can be a gene locus on the Salmonella chromosome by designing a homologous fusion fragment of the target gene, it is cloned into a suicide vector, and the suicide vector is transformed into the target bacterium. An insertion mutant is selected by antibiotic screening. Under the second round of reverse selection pressure, only the mutation that contain second homologous recombination and the loss of the suicide plasmid can survive. By PCR screening and sequencing, we can obtain the mutant of the strain.
Fig.1. Workflow of homologous recombination genome editing
• Homology arms design and suicide plasmid construction
CRISPR/Cas9-mediated Salmonella Genome Editing
CRISPR technology, which derived from the immune system present in bacteria and archaea, is an efficient genome-scale editing tool that has revolutionized conventional genetic engineering methods and unprecedentedly facilitated strain engineering. It enables fast and reliable genetic manipulation in Salmonella. Two components are requested to work: a guide RNA (gRNA), e.g. under an RNA polymerase III promoter, and the nuclear localization tag fused DNA endonuclease, with Cas9 being the most commonly used.
Here we use CRISPR/Cas9 machinery coupled to lambda (λ) recombinase-mediated homologous recombination (recombineering) to accomplish the Salmonella genome editing.
When Cas9 protein and gRNA are expressed in bacteria cells, Cas9 introduces DSBs that must be repaired by the cells via non-homologous end joining (NHEJ) or homologous recombination (HR). By supplying a DNA repair template for use in HR, various DNA modifications can be obtained.
Fig.2. Workflow of CRISPR /Cas9-mediated Salmonella Genome Editing
• sgRNA design and construction
The advantages of CRISPR based Salmonella genome editing in Microbiosci:
• Fast turnaround time
• Scarless genome editing
• Multigene editing: can knock-out up to 5 genes simultaneously
• Easy selection: no selectable marker is required
What we could help?
• Gene disruption, deletion or insertion
• Reporter gene and tag integration
• Promoter fine tuning
• Introduction of point mutations
With years of experience in genome editing field, Microbiosci could provide the most excellent service for Salmonella genome editing. Our talent experts are dedicated to edit your Salmonella genome with the greatest chance to succeed. Meanwhile, based on the commitment of prompt communication and on-time reporting, our staffs will ensure a high-efficiency service to meet the strict project timelines.
If you have any special requirements in our Salmonella genome editing service, please feel free to contact us at firstname.lastname@example.org or 1-631-626-9181. We are looking forward to working together with your attractive projects.
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