Cas9 Lentivirus (Puromycin Selection)

Catalog #

78066

$795 *

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 Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed 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 Cas9 gene driven by an EF1A promoter, along with a Puromycin selection marker.

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

Figure 1: Schematic of the lenti-vector used to generate the Cas9 Lentivirus.

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Synonyms

Cas9 Lentivirus, CRISPR lentivirus, spCas9 lentivirus

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Product Data Gallery

Cas9 expression in A549 cells transduced with Cas9 Lentivirus (Puromycin Selection) (BPS Bioscience #78066).

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 lentivirus particles were produced in HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.

Background

Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by a sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the targeted gene. Cas9 Lentivirus can be used to generate Cas9 expressing cells in almost any mammalian cell line. Cells stably expressing Cas9 can then be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines. The use of Cas9 technology has revolutionized the cell biology field, and its power can be harnessed for therapeutical applications.

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

Expression of Cas9 in target cells
Generation of a stable Cas9 over-expressing cell line following puromycin 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

CRISPR

Regulatory Status

Research Use Only

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Cas9 Lentivirus (Puromycin Selection)

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