MART-1-Specific TCR Lentivirus (Clone DMF4)

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78678

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Description

The human MART-1-specific T Cell Receptor lentiviruses (clone DMF4) 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. These viruses transduce a human TCR (clone DMF4) that specifically recognizes antigen MART-1 (Melanoma-associated antigen recognized by T cells-1, or Melan-A), and in which the TCR α chain and β chain are linked by P2A (Figure 1). The lentiviruses also transduce a puromycin selection gene.

Figure 1. (A) Schematic of the lenti-vector used to generate the MART-1-specific TCR lentivirus and (B) Construct diagram showing expressed components of the MART-1-specific TCR. TRAV and TRAC correspond to the TCR alpha chain variable and constant regions, respectively, whereas TRBV and TRBC correspond to the TCR beta chain variable and constant regions.

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Product Data Gallery

Expression of MART-1-specific TCR in Jurkat cells transduced with lentiviruses.

Expression of MART-1-specific TCR (clone DMF4) in TCR knockout NFAT Luciferase Reporter Cell Line confers responsiveness to anti-CD3 agonist and induces NFAT-dependent luciferase activity.

T Cell Activation using T2 cells as APC.

T Cell Activation using A375 cells as APC.

Product Info

Storage and Usage

Citations

Supplied As

≥2x10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
Thaw Medium 2	BPS Bioscience #60184
Growth Medium 2C	BPS Bioscience #79592
Assay Medium 2D	BPS Bioscience #78755
TCR KO NFAT Luciferase Reporter Jurkat Cell Line	BPS Bioscience #78556
CD8⁺ TCR KO NFAT Luciferase Reporter Jurkat Cell Line	BPS Bioscience #78757
T2 Cell Line	ATCC #CRL-1992
A375 Cells	ATCC #CRL-1619
NY-ESO-1 (157-165) Peptide	BPS Bioscience #78758
MART-1 (26-35) Peptide	BPS Bioscience #78759
MART-1 (Leu26-35, Leu27) Peptide	BPS Bioscience #78760
MART-1 (27-35) Peptide	BPS Bioscience #78761
APC MHC I Dextramer (HLA-A*02:01 SLLMWITQV)	Immudex #WB03247
PE anti-human α/β T Cell Receptor Antibody	BioLegend #306707
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Polybrene	Millipore Sigma #TR-1003-G

Formulation

The lentiviruses were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee

Background

MART-1 is a differentiation antigen expressed at the surface of melanocytes. A peptide fragment of the protein is found bound to MHC complexes that are recognized by cytotoxic T cells. MART-1 is found in most skin cancers, including melanomas, and is used as a biomarker for diagnostic purposes. Since the expression of the protein is restricted to melanocyte containing tissues (skin and retina), and is not found in other tissues, MART-1 is an attractive target for cancer vaccines and adoptive cell therapy. MART-1 peptide 26-35 is a fragment commonly associated with MHC and recognized by T cell receptors.

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.

Growth Media

For best results, the use of BPS Bioscience validated and optimized media is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for Maintaining CD8⁺ TCR KO NFAT Luciferase Reporter Cell Line
Growth Medium 2C (BPS Bioscience #79592): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 1 mg/ml Geneticin, 100 µg/ml Hygromycin B

Media Required for Maintaining T2 Cells
Thaw Medium 2 (BPS Bioscience #60184): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin

Media Required for Co-culture Assay
Assay Medium 2D (BPS Bioscience #78755): RPMI 1640 medium supplemented with 1% FBS

Applications

Use as a positive control for MART-1 TCR evaluation and optimize experimental conditions.
Generate MART-1 TCR stable cell line (puromycin resistant).

Shipping Temperature

-80°C

Notes

To generate stable TCR-expressing cells, remove the growth medium 48-72 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin for antibiotic selection of transduced cells. For more information on how to determine the concentration of antibiotic required, see our FAQs resources “what is a kill curve?”

Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and 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. 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.

License Disclosure
Visit bpsbioscience.com/license for the label license and other key information about this product.

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

CAR T-Cell Therapy

Regulatory Status

Research Use Only

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MART-1-Specific TCR Lentivirus (Clone DMF4)

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