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: Lymphocyte-activation gene 3 CRISPR, CD223 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: Lymphocyte-activation gene 3 (LAG3, CD223) is a cell surface protein that belongs to the immunoglobulin (Ig) superfamily. LAG3 is expressed on activated T-cells, Natural Killer cells, B-cells, and plasmacytoid dendritic cells. Its main ligand is the MHC class II, to which it binds with higher affinity than CD4. It negatively regulates cellular proliferation, activation, and homeostasis of T-cells in a similar fashion as CTLA-4 and PD-1, and has been reported to play a role in T-reg suppressive function. A number of LAG3 antibodies are in preclinical development for the treatment of cancer and autoimmune disorders. LAG3 may be a better immune checkpoint inhibitor target than CTLA-4 or PD-1, because antibodies targeting CTLA-4 or PD-1 only activate effector T-cells while failing to inhibit T-reg activity, whereas an antagonist LAG3 antibody can both activate effector T-cells (by downregulating the LAG3 inhibiting signal) and inhibit induced (i.e. antigen-specific) T-reg suppressive activity.The LAG3 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 LAG3 (GenBank Accession #NM_002286) driven by a U6 promoter. Note: unlike Human LAG3 CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78053), the Human LAG3 CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, resulting in only transient expression of the Cas9 and LAG3-targeting sgRNA. While this may minimize potential off-targeting risks due to either prolonged expression or integration of the Cas9, puromycin selection should not be used for more than 48 hours post-transduction, which may lower knockout efficiency.
• 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 LAG3 in a target cell pool.2. Generation of a stable LAG3 knock-out cell line following limited dilution.
• Product Type: Lentivirus
• Biosafety Level: BSL-2
• Related Products: 71213, 79813, 78052, 78053
• 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.