CD5 (Human) CRISPR/Cas9 Lentivirus (Non-Integrating)

Catalog #

78198

$995 *

Size: 500 µl x 2

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

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Description

CD5 is a member of the scavenger receptor cysteine-rich (SRCR) superfamily and is associated with the immune system. This protein is a transmembrane glycoprotein found on the surface of thymocytes and may act as a receptor to regulate T-cell proliferation.

The CD5 CRISPR Lentiviruses are replication incompetent, HIV-based VSV-G pseudotyped 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 5 sgRNA (single guide RNA) targeting human CD5 driven by a U6 promoter (Figures 1 and 2).

Unlike CD5 CRISPR/Cas9 Lentivirus (Integrating) (BPS Bioscience, #78119), the CD5 (Human) CRISPR/Cas9 Lentivirus (Non-Integrating) is made with a mutated Integrase, therefore the viral DNA cannot integrate into the genome, resulting in only transient expression of the Cas9 and CD5-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. Efficiencies may vary, depending on the cell type and the gene of interest.

Figure 1. Schematic of the Lenti-vector used to generate the CD5 CRISPR/Cas9 Lentivirus

Gene Target:	Primer ID:	sgRNA Sequence
CD5	CD5-1	CAGCATCTGTGAAGGCACCG
CD5	CD5-2	TTTCCTGAAGCAATGCTCCA
CD5	CD5-3	AAGCGTCAAAAGTCTGCCAG
CD5	CD5-4	GCTGTAGAACTCCACCACGC
CD5	CD5-5	CGTTCCAACTCGAAGTGCCA

Figure 2. List of sgRNA Sequences in the CD5 CRISPR/Cas9 Lentivirus

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Synonyms

CD5 molecule, T1, LEU1, CD5 protein, CD5 scavenger receptor cysteine-rich superfamily (SRCR), CD5 lentivirus, CD5 CRISPR

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Product Data Gallery

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 CD5 in target cells.
2. Generation of stable CD5 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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CD5 (Human) CRISPR/Cas9 Lentivirus (Non-Integrating)

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