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Gene Editing and Virus Facility

Cutting-edge genetic manipulation and viral vector production

Services

Our unit operates on a complete-service fully costed model, tailoring our charges based on researcher affiliation.

For advice on using CRISPR/Cas9 in your research or should you require a novel transgenic mouse or cell line, please email: gevf@embl.it

For advice on using viruses for your research or ordering a virus, please email: viruses@embl.it

Mouse and cell service elements include:

  • CRISPR/Cas9 project design and reagent sourcing
  • Zygote pronuclear microinjection
  • Surgical embryo implantation
  • Microinjection of targeted ES cells into host 8-cell embryos
  • Sequence confirmation of F1 genotype
  • Cryopreservation of embryos and/or sperm
    IVF rederivation
  • Import and rederivation of mouse lines into the LAR facility

Virus service elements include:

Production of custom-made purified and quantitated viral stocks that are ready for cargo delivery to the desired cell type.

Recombinant Adeno-Associated Virus (rAAV)

  • Vector capacity: 4.7Kb
  • Cargo remains episomal
  • Current serotypes in stock include rAAV1, 2, 1/2 chimeric, 2-retro, DJ, DJ/8, DJ/9, 5, 6, 7, 8, 8BP2, 9, PHP.S, PHP.EB, B10, and 7m8. 
  • Please let us know if you require another serotype and we will do what we can to obtain the necessary reagents.
  • Typically provide at least 200μl of ≥1×1012 VG/ml but smaller or larger quantities can be provided at request.

Lentivirus

  • Vector capacity: 8Kb
  • Cargo integrates
  • Currently, we utilise VSV-G due to its wide range of infectivity. Other envelope proteins could be offered if requested. Currently, viruses are produced using either the 2nd or 3rd generation.

Herpes Simplex Virus (HSV)

  • Vector capacity: 30Kb
  • Cargo remains episomal

DNA constructs

  • Long ssDNA (up to ~5kb) for CRISPR/Cas9-mediated knock-in
  • Modification of DNA, precise to the base pair
  • Targeting construct to make a knock-in or knockout
  • General sub-cloning service
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