BCMA CRISPR/Cas9 Lentivirus (Integrating)

Catalog #

78893

$835 *

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

The BCMA CRISPR/Ca9 Lentiviruses are replication incompetent, HIV-based, VSV-G pseudo-typed lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses contain a CRISPR/Cas9 gene, driven by an EF1A promoter, and 5 sgRNA (single guide RNA) targeting human BCMA (B-cell maturation antigen) driven by a U6 promoter (see Table 1 for sgRNA sequences). The integrating lentivirus integrates randomly into the cell’s genome to express both the Cas9 and sgRNAs. Simultaneous expression of Cas9 and BCMA sgRNA allows the generation of BCMA-knockout cells with one single transduction step. The lentiviruses also contain a puromycin selection marker (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate the BCMA CRISPR/Cas9 Lentivirus.

Table 1: List of sgRNA sequences in the BCMA CRISPR/Cas9 Lentivirus.

Gene Target:	Primer ID:	sgRNA Sequence:
TNFRSF17 (BCMA)	TNFRSF17-1	CCTCTAACATGTCAGCGTTA
TNFRSF17 (BCMA)	TNFRSF17-2	TGTCAACTTCGATGTTCTTC
TNFRSF17 (BCMA)	TNFRSF17-3	CGAGTACACGGTGGAAGAAT
TNFRSF17 (BCMA)	TNFRSF17-4	TTCACTGAATTGGTCACACC
TNFRSF17 (BCMA)	TNFRSF17-5	GTGTTTTTAAACTCGTCCTT

Note: BPS Bioscience also offers a non-integrating version of this product (BPS Bioscience #78894).

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Synonyms

knockout, knock-out, KO

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Product Data Gallery

Flow cytometry analysis of BCMA expression in RPMI-8226 cells transduced with BCMA CRISPR/Cas9 lentivirus

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥1 x 10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.      

Background

B-cell maturation antigen (BCMA), also known as CD269 or tumor necrosis factor receptor superfamily member 17 (TNFRSF17), is a cell surface receptor of the TNF receptor superfamily that recognizes B-cell activating factor (BAFF) and is involved in B cell proliferation and maturation. BCMA is preferentially expressed in mature B lymphocytes and the soluble form of BCMA can be found at higher levels in the serum of Multiple Myeloma (MM) patients. BCMA is a highly attractive target antigen for immunotherapy. BCMA, similarly to CD19, is restricted in expression to mature B cells, allowing the progenitor’s population to be spared during treatment and to replenish the patient’s B cell population. BMCA targeting therapies include bispecific antibodies, antibody-drug conjugates, and chimeric antigen receptor (CAR) T cells. To date, the FDA has approved two BCMA CAR-T therapies for the treatment of MM, that resulted in promising outcomes for patients. Further studies will allow a better understanding of the role of BCMA in cancer and fine tune cancer therapy tools.

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.

Applications

Knock-out of BCMA in cells of interest.
Generation of BMCA knock-out cell pools.
Generation of a stable BCMA knock-out cell line following puromycin selection and single-cell selection.

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.

Biosafety
The lentiviruses are produced with the SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Immunotherapy Immune Checkpoints

Regulatory Status

Research Use Only

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BCMA CRISPR/Cas9 Lentivirus (Integrating)

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