CRBN CRISPR/Cas9 Lentivirus (Non-Integrating)

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Applications: Transient knockdown of CRBN in target cell poolsGeneration of stable CRBN knockout cells following transient puromycin selection (48h maximum) followed by limiting dilution cloning.

Description: Cereblon (CRBN) forms an E3 ubiquitin ligase complex which is responsible for ubiquitinating proteins that regulate various developmental processes. CRBN also binds to Calcium Activated Potassium Channel subunit alpha-1 (KCNMA1) to regulate ion transport. Moreover, mutations in CRBN may play an underlying role in tumor cells acquiring resistance to immunotherapy.The CRBN CRISPR/Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed lentiviral particles that are ready to transduce 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 5 sgRNA (single guide RNA) targeting human CRBN.The non-integrating lentivirus is made with a mutated integrase, resulting in only transient expression of the Cas9 and sgRNA. Although using the non-integrating lentivirus results in lower knockdown efficiency, the Cas9 is not permanently expressed, which lowers the risk of off-targeting, and there are no random integrations into the cell's genome. Knockout cell lines can still be generated following cell sorting or limited dilution, because even though the Cas9 and sgRNA expression is transient, the changes in the genomic DNA from the Cas9 nuclease activity and NHEJ repair are permanent.

Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture 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 10⁷ TU/ml. The titer will vary with each lot; the exact value will be provided with each shipment.

Warnings: Avoid freeze/thaw cycles.

Biosafety Level: BSL-2

• Artikelnummer: BPS78518
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 78518
• Verpackungseinheit: 500 µl x 2
• Mengeneinheit: STK