DESCRIPTION

• Notes: The CRISPR/CAS9 technology is covered under numerous patents, including U.S. Patent Nos. 8,697,359 and 8,771,945, as well as corresponding foreign patents applications, and patent rights.
• Species: Replication incompetent HIV
• Shiptemp: -80°C (dry ice)
• Synonyms: Programmed Cell Death Protein 1 CRISPR, PD-1 lentivirus
• Warnings: Biosafety: None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells. Although the pseudotyped lentiviruses are replication-incompetent, they do require the use of a Biosafety Level 2 facility. BPS recommends following all federal, state, local, and institutional regulations and using all appropriate safety precautions.
• Category: CRISPR/Lentivirus
• Description: The binding of Programmed Cell Death Protein 1 (PD-1), a receptor expressed on activated T-cells, to its ligands, PD-L1 and PD-L2, negatively regulates immune responses. The PD-1 ligands are found on most cancers, and the PD-1:PD-L1/2 interaction inhibits T-cell activity and allows cancer cells to escape immune surveillance. The PD-1:PD-L1/2 pathway is also involved in regulating autoimmune responses, making these proteins promising therapeutic targets for a number of cancers, as well as multiple sclerosis, arthritis, lupus, and type I diabetes. The PD-L1 CRISPR Lentiviruses are replication incompetent, HIV-based, VSV-G pseudo-typed lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a CRISPR/Cas9 gene driven by an EF1a promoter, along with 4 sgRNA (single guide RNA) targeting human PD-L1 (Programmed Cell Death 1 Ligand 1, CD274, B7 homolog 1 (B7-H1), GenBank accession #NM_021893) driven by a U6 promoter (Figures 1 and 2). The integrating lentivirus integrates randomly into the cell's genome to express both the Cas9 and sgRNA. Puromycin selection increases the knockout efficiency by forcing high expression levels of both Cas9 and the sgRNA, and can be used with the integrating lentivirus to quickly and easily achieve high knockdown efficiencies in a cell pool. Efficiencies also depend on the cell type and the gene of interest.
• Formulation: The lentiviruses were produced from HEK293T cells in medium containing 90% DMEM + 10% FBS.
• Supplied As: Two vials (500 µl x 2) of lentivirus at a titer ≥1 x 10^6 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
• Unspsc Code: 41106621
• Unspsc Name: Virus mediated expression vectors or kits
• Applications: 1. Transient knock-down of PD-L1 in target cells.2. Generation of a stable PD-L1 knock-out cell line following puromycin selection.
• Product Type: Lentivirus
• Biosafety Level: BSL-2
• Related Products: 60543, 78052, 78055, 78064
• Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
• Scientific Category: Immunotherapy
• Instructions for Use: See assay protocol for detailed instructions.