CD47 CRISPR/Cas9 Lentivirus (Non-Integrating)

Catalog #

78063

$995 *

Size: 500 µl x 2

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

CD47 (also known as Rh-associated protein, GP42, Integrin-Associated Protein (IAP), or Neurophilin) is an immunoglobulin-like protein that interacts with its receptor, Signal-regulatory protein alpha (SIRPα), on macrophages. This binding interaction regulates transmigration, oxidative burst cytokine production, and phagocytosis, generating a “don’t eat me” signal. CD47 is ubiquitously expressed on the surface of normal cells, but is overexpressed in numerous cancer cells where it is thought to contribute to the resistance of tumors to phagocyte-dependent clearance.

The CD47 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 CD47 (NM_198793.2) 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 CD47 CRISPR/Cas9 Lentivirus.

Gene Target:	Primer ID:	sgRNA Sequence
CD47	CD47-1	ATCGAGCTAAAATATCGTGT
CD47	CD47-2	GCACTTAAATATAGATCCGG
CD47	CD47-3	AGTGATGCTGTCTCACACAC
CD47	CD47-4	TTTGCACTACTAAAGTCAGT

Figure 2. List of sgRNA Sequences in the CD47 CRISPR/Cas9 Lentivirus.

Note: unlike human CD47 CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78056), the human CD47 CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, resulting in only transient expression of the Cas9 and CD47 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

Rh-associated protein CRISPR, GP42 lentivirus, Integrin-Associated Protein (IAP) CRISPR, Neurophilin lentivirus

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Product Info

Storage and Usage

Citations

Species

Replication Incompetent HIV

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 CD47 in a target cell pool.
2. Generation of stable CD47 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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CD47 CRISPR/Cas9 Lentivirus (Non-Integrating)

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