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Applications: Express SaCas9 to enable genome editing
Description: Adeno-Associated Virus serotype 9 (AAV9) is one of the most promising serotypes for gene therapy applications. AAV9 transduces a wide range of tissue types, including cardiac and skeletal muscle, and liver, pancreas, and eye tissue. AAV8 and AAV9 have recently been used to correct disease-causing mutations and improve muscle function in mouse models of Duchenne muscular dystrophy. AAV9 has significantly lower seroprevalence in the human population than other AAV serotypes, making AAV9 a desirable candidate for therapeutic applications.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.
Formulation: AAV was produced in HEK293-AAV cells and is supplied in PBS-MK (PBS Magnesium-Potassium) buffer containing 0.01% Pluronic F68.
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.
Storage Stability: AAV is shipped with dry ice. For long-term storage, it is recommended to store AAV at -80°C.
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.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
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