PD-L1 CRISPR/Cas9 Lentivirus (Non-Integrating)

Catalog #

78064

$995 *

Size: 500 µl x 2

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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).

Figure 1. Schematic of the Lenti-vector used to generate the TIGIT CRISPR/Cas9 Lentivirus.

Figure 1. Schematic of the Lenti-vector used to generate the PD-L1 CRISPR/Cas9 Lentivirus.

Gene Target:	Primer ID:	sgRNA Sequence
PD-L1	PD-L1-1	GGTTCCCAAGGACCTATATG
PD-L1	PD-L1-2	ACTGCTTGTCCAGATGACTT
PD-L1	PD-L1-3	GCATAGTAGCTACAGACAGA
PD-L1	PD-L1-4	ACATGTCAGTTCATGTTCAG

Figure 2. List of sgRNA Sequences in the PD-L1 CRISPR/Cas9 Lentivirus.

Note: unlike Human PD-L1 CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78057), the Human PD-L1 CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, resulting in only transient expression of the Cas9 and PD-L1-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.

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Synonyms

Programmed Cell Death Protein 1 CRISPR, PD-1 lentivirus

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Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 μl x 2) of lentivirus at a titer ≥1 x 10⁶ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

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 lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.

Instructions for Use

See assay protocol for detailed instructions.

Applications

1. Transient knock-down of PD-L1 in a target cell pool.
2. Generation of stable PD-L1 knock-out cells using transient puromycin selection (48h maximum) followed by limited dilution.

Shipping Temperature

-80°C

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.

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.

Scientific Category

Immunotherapy

Regulatory Status

Research Use Only

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PD-L1 CRISPR/Cas9 Lentivirus (Non-Integrating)

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