Creative Biogene is a leading biotechnology company offering the best Staphylococcus aureus 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 Staphylococcus aureus genome editing services.
Our Staphylococcus aureus genome editing services based on CRISPR/Cas9 technology and homologous recombination technique. The state-of-art Staphylococcus aureus editing system helps you successfully achieve gene knockout, gene insertion and point mutation for either research or industrial purposes.
Staphylococcus aureus Genome Editing Based on Homologous Recombination
Red/ET Recombination permits the engineering of DNA in LAB 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 LAB 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 Staphylococcus aureus. 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
• Even the most demanding tasks can be reduced to four basic steps:
1. Homology arms design and suicide plasmid construction
CRISPR /Cas9-mediated Staphylococcus aureus 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 Staphylococcus aureus. 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 Staphylococcus aureus genome editing.
When Cas9 protein and gRNA are expressed in bacteria cells, Cas9 introduces DSBs that must be repaired by the cells via homologous recombination (HR). By supplying a DNA repair template for use in HR, various DNA modifications can be obtained.
Fig.2. Diagram of the CRISPR/Cas9-mediated genome editing
• Project steps:
1. sgRNA design and construction
• The advantages of CRISPR based Staphylococcus aureus genome editing in Creative Biogene:
1. Fast turnaround time
2. Scarless genome editing
3. Multigene editing: can knock-out up to 5 genes simultaneously
4. 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, Creative Biogene could provide the most excellent service for Staphylococcus aureus genome editing. Our talent experts are dedicated to edit your Staphylococcus aureus 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.
1.Monk, I. R. . (2012). Genetic manipulation of staphylococci—breaking through the barrier. Frontiers in Cellular and Infection Microbiology, 2.
2. Liu, Q., Jiang, Y., Shao, L., Yang, P., Sun, B., Yang, S., & Chen, D. (2017). CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus. Acta Biochimica et Biophysica Sinica, 49(9), 764–770.