CBL-B (Human) CRISPR/Cas9 Lentivirus (Integrating)

Catalog #

78343

$795 *

Size: 500 µl x 2

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Description

CBL-B is an E3 ubiquitin-protein ligase which has been identified as a negative regulator of T-cell activation. Using CRISPR/Cas9 to inactivate CBL-B has been shown to be sufficient to inhibit T-cell expansion.

The CBL-B CRISPR/Cas9 Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to infect 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 5 sgRNA (single guide RNAs) targeting human CBL-B driven by a U6 promoter (Figures 1 and 2).

The integrating lentivirus integrates randomly into the cellular genome to express both Cas9 and the sgRNA. Puromycin selection forces high expression levels of both Cas9 and the sgRNA, and can be used with the integrating lentivirus to quickly and easily achieve high knockdown efficiencies in a cell pool. Efficiencies may vary, depending on the cell type and the gene of interest.

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

Gene Target:	sgRNA Sequence
CBL-B	TGTGGGATGTCGACTCCTAG
CBL-B	CTTCATCTCTTGGATCAAAG
CBL-B	TTCCGCAAAATAGAGCCCCA
CBL-B	TGAATTAGATCCAGGCGAGG
CBL-B	TGCACAGAACTATCGTACCA

Figure 2. List of sgRNA Sequences in the CBL-B CRISPR/Cas9 Lentivirus

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Synonyms

CBL-B, CBLB, Cbl proto-oncogene B, RING finger protein 56, RNF56, Nbla00127, E3 ubiquitin-protein ligase CBL-B, Casitas B-lineage lymphoma proto-oncogene b

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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.

Applications

1. Transient knock-down of CBL-B in target cells
2. Generation of a stable CBL-B knock-out cell pool following puromycin selection and 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.

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

Warnings

Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Biosafety
None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Scientific Category

CRISPR

Regulatory Status

Research Use Only

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CBL-B (Human) CRISPR/Cas9 Lentivirus (Integrating)

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