TIGIT CRISPR/Cas9 Lentivirus (Non-Integrating)

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Applications: 1. Transient knock-down of TIGIT in a target cell pool.
2. Generation of a stable TIGIT knock-out cell line following limited dilution.

Description: TIGIT (T-cell immunoreceptor with Ig and ITIM domains; VSTM3; VSIG9) is a co-inhibitory receptor that is highly expressed in Natural Killer (NK) cells and activated CD4+, CD8+, and regulatory T-cells. Interaction with the Poliovirus Receptor (PVR; CD155) on antigen presenting cells, such as dendritic cells, recruits either the Src homology (SH) domain-containing tyrosine phosphatases SHP1 and SHP2, or the Inositol phosphatase SHIP1 and SHIP2, to the TIGIT ITIM domain. This increases IL-10 release and suppresses NF-κB and NFAT T-cell receptor (TCR) signaling, which blocks T-cell proliferation and cytokine production. TIGIT also serves as a competitive inhibitor of CD226, a costimulatory receptor for CD155. TIGIT-targeting antibodies which block this T-cell intrinsic inhibitory effect have shown enhanced anti-tumor and anti-viral functions in preclinical studies.
The TIGIT 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 TIGIT (GenBank Accession #NM_173799) driven by a U6 promoter (Figures 1 and 2).Note: unlike human TIGIT CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78058), the human TIGIT CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, resulting in only transient expression of the Cas9 and TIGIT 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.

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.

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.

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.

Biosafety Level: BSL-2

• Artikelnummer: BPS78065
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 78065
• Verpackungseinheit: 500 µl x 2
• Mengeneinheit: STK