B2M (Human) CRISPR/Cas9 Lentivirus (Non-Integrating)

Catalog #

78341

$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

Beta-2 Microglobulin (B2M) is a required component of Major Histocompatibility Complex (MHC) class 1 molecules, which present peptide fragments from within the cell to cytotoxic T-cells as part of the adaptive immune system.

The B2M 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 B2M driven by a U6 promoter (Figures 1 and 2).

Note: unlike B2M CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78340), the B2M CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, resulting in only transient expression of the Cas9 and B2M-targeting sgRNA.

It is expected that this will minimize potential off-target effects caused by either prolonged expression or random integration of Cas9 and the sgRNA. A short round of puromycin selection right after transduction may increase knockout efficiency, however puromycin should not be used for more than 48 hours post-transduction due to the transient nature of expression using the non-integrating lentivirus.

B2M (Human) CRISPR/Cas9 Lentivirus

Figure 1. Schematic of the Lenti-vector used to generate the B2M CRISPR/Cas9 Lentivirus

Gene Target	sgRNA Sequence
B2M	AAGTCAACTTCAATGTCGGA
B2M	CTGAATCTTTGGAGTACCTG
B2M	GAGTAGCGCGAGCACAGCTA
B2M	TCCTGAATTGCTATGTGTCT
B2M	GAAGTTGACTTACTGAAGAA

Figure 2. List of sgRNA Sequences in the B2M CRISPR/Cas9 Lentivirus

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Synonyms

B2M, beta-2-microglobulin, IMD43

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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 B2M in target cells
2. Generation of stable B2M 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.

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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B2M (Human) CRISPR/Cas9 Lentivirus (Non-Integrating)

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