HLA-E Lentivirus

Catalog #

78929

$1,250 *

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

HLA-E Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses result in expression of the human HLA-E heavy chain (NM_005516.6) driven by an EF1a promoter, and a puromycin selection marker (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate HLA-E Lentivirus (encoding HLA-E heavy chain).

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Expression of HLA-E in K562 cells transduced with HLA-E Lentivirus

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥10⁷ 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 in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.

Background

HLA-E, or MHC (major histocompatibility complex) class I antigen E, is considered a non-classical MHC class I with low expression and fewer polymorphisms than the remaining HLA. HLA-E is composed of a heavy chain and β-2 microglobulin (B2M). It binds to specific peptides derived from the classical MHC class I (HLA-A, B, C and G), after these have been processed in the endoplasmic reticulum and the proteosome. The complex of HLA-E with the peptide is recognized by NK cells via the inhibitory receptor CD94/NKG2A/B. Binding to CD94/NKG2C however results in NK cell activation. Expression of HLA-E combined with knockout of HLA-A, B and C, in pluripotent stem cell (PSC) and their differentiated cell types, resulted in these cells escaping attack by CD8⁺ T cells and NK cytotoxicity. This strategy brings us closer to an almost universal cell donor reality, reducing the risk of immune rejection during cell transplants and alleviating the enormous investment of creating a PSC bank that has representation of all the haplotypes.

References

Gornalusse G., et al., 2017 Nature Biotechnology 35:765-772

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Applications

Expression of human HLA-E in cells of interest.
Generate cell pools or stable cell lines expressing HLA-E following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate an HLA-E stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection.

Biosafety
The lentiviruses are produced with a 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 further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

CAR T-Cell Therapy

Regulatory Status

Research Use Only

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HLA-E Lentivirus

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