Dominant Negative TGF-β Receptor Type II (TGF-βRII) Lentivirus

Catalog #

78928

$1,250 *

Size: 500 µl x 2

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Description

Dominant Negative TGF-β Receptor Type II (TGF-βRII) 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 human dominant negative TGF-βRII, missing the intracellular kinase domain (NM_003242.6; amino acid 1-191), driven by an EF1a promoter and a puromycin selection marker (Figure 1).

Figure 1. Schematic of the lenti-vector used to generate Dominant Negative TGF-β Receptor Type II (TGF-βRII) Lentivirus

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Synonyms

TGFb-1, TGFb1

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Product Data Gallery

Expression of dominant negative TGF-βRII in CHO cells transduced with Dominant Negative TGF-β Receptor Type II (TGF-βRII) 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

Transforming growth factor receptor beta 2 (TGFBR2), or TGFβRII, encodes the TGFβ receptor serine/threonine kinase, which is a transmembrane protein that forms a heterodimeric complex with other receptor proteins and binds TGFβ. The association of TGFβ with TGFβRII leads to the phosphorylation of proteins, such as SMADs, involved in cell proliferation, cell cycle arrest, wound healing, and immunosuppression. Dysfunction of the TFGβ signaling tends to result in cancer development and progression. In the case of solid tumors, TGFβ signaling plays a role in creating a highly immunosuppressive TME (tumor microenvironment), restricting the efficacy of CAR (chimeric receptor antigen)- T cells, which have proved successful in the treatment of hematological cancers. Recently, the use of CAR-T cells armored with a dominant negative form of TGFβRII, missing the intracellular kinase domain, for the treatment of prostate cancer resulted in promising outcomes. 5 out of 13 patients did suffer cytokine release syndrome, which continues to be a concern with CAR-T applications, but on the whole the use of a dominant negative TGFβ receptor to armor CAR-T cells appears to be an approach that deserves further attention in the cancer therapy field.

References

Ikushima H. and Miyazono K., 2010 Nature Reviews Cancer 10:415-424.
Narayan V., et al., 2022 Nat Med 28:724-34.

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 dominant negative TGF-βRII in cells of interest.
Generate cell pools or stable cell lines expressing human TGF-βRII following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a dominant negative TGF-βRII expressing 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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Dominant Negative TGF-β Receptor Type II (TGF-βRII) Lentivirus

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