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BPS Bioscience

DETAILS

  • Notes: Biosafety: Recombinant AAV is inherently replication-deficient and not known to cause any human diseases. Additionally, following transduction, AAV vectors exist episomally and do not integrate into or disrupt the host cell's genome. AAV requires the use of a Biosafety Level 1 facility. BPS Bioscience recommends following all local, federal, state, and institutional regulations and using all appropriate safety precautions. Troubleshooting Guide: Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email support@bpsbioscience.com. Licensing Agreement: The AAV-DJ viruses are covered under several patents, including U.S. Patent Nos. 7,588,772, 8,067,014, 8,574,583, and 8,906,387, as well as corresponding foreign patents applications and patent rights. AAV-DJ is used under a license agreement.
  • Species: AAV-DJ
  • Shiptemp: -80°C (dry ice)
  • Warnings: Avoid freeze/thaw cycles
  • Category: Viral Tools/AAV
  • Description: Cas9 is an endonuclease enzyme that introduces a double stranded break into the DNA when recruited to a specific DNA sequence by the sgRNA (single guide RNA). This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the gene. SaCas9 (Staphylococcus aureus CRISPR-associated protein 9) has demonstrated high cutting efficiency in mammalian cells, and its smaller size makes it ideal for packaging into AAV. SaCas9 recognizes a longer protospacer adjacent motif (PAM) site, 5'-NNGRRT-3', than the more traditional SpCas9 (Streptococcus pyogenes CRISPR-associated protein 9). These AAV particles constitutively express SaCas9 under the control of a CMV promoter. Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling. These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types.
  • Formulation: AAV-DJ was produced in HEK293-AAV cells and is supplied in PBS-MK (PBS Magnesium-Potassium) buffer containing 0.01% Pluronic F68.
  • Supplied As: Two vials (50 µl x 2) of AAV at a titer ≥1 x 1012 TU/ml. The titer is determined by qPCR and will vary with each lot; the exact value will be provided with each shipment.
  • Unspsc Code: 41106621
  • Unspsc Name: Virus mediated expression vectors or kits
  • Applications: Generate a SaCas9 over-expressing cell line for knockout or knock-in studies.
  • Product Type: AAV virus
  • Purification: The purity of the AAV particles was confirmed to be greater than 90% by staining with One-Step Lumitein™ UV Protein Gel Stain (Biotium #21005-1L). The purity will vary with each lot; the exact value will be provided with each shipment.
  • Biosafety Level: BSL-1
  • Related Products: 78443, 78463, 78469, 78442
  • Storage Stability: AAV is shipped with dry ice. For long-term storage, it is recommended to store AAV at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
  • Scientific Category: Viral Tools
  • Signed License Required: No

DESCRIPTION

Cas9 is an endonuclease enzyme that introduces a double stranded break into the DNA when recruited to a specific DNA sequence by the sgRNA (single guide RNA). This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the gene. SaCas9 (Staphylococcus aureus CRISPR-associated protein 9) has demonstrated high cutting efficiency in mammalian cells, and its smaller size makes it ideal for packaging into AAV. SaCas9 recognizes a longer protospacer adjacent motif (PAM) site, 5'-NNGRRT-3', than the more traditional SpCas9 (Streptococcus pyogenes CRISPR-associated protein 9). These AAV particles constitutively express SaCas9 under the control of a CMV promoter. Adeno-Associated Virus-DJ (AAV-DJ) is a synthetic serotype made from eight different wild-type AAV serotypes (AAV2, 4, 5, 8, 9, avian, bovine, and goat AAV) using DNA shuffling. These modifications give the AAV-DJ serotype improved transduction efficiency in vitro and in vivo compared to wild-type serotypes. Consequently, AAV-DJ can infect a broad range of cell types.