CD8+ TCR Knockout NFAT-Luciferase Reporter Jurkat Cell Line

Catalog #

78757

$16,000 *

Size: 2 vials

Qty

Bulk/Custom

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

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Description

This cell line was generated from TCR Knockout NFAT Luciferase Reporter Jurkat Cell Line (BPS Bioscience #78556) by exogeneous overexpression of human CD8 (NM_001768.6) using lentiviral transduction (CD8a Lentivirus, BPS Bioscience #78648). To achieve knockout of TCR (T Cell Receptor), the TRAC (T-Cell Receptor Alpha Constant) and TRBC1 (T-Cell Receptor Beta Constant 1) domains of the TCRα/β chains were genetically removed by CRISPR/Cas9 genome editing from recombinant Jurkat cells stably expressing the firefly luciferase gene under the control of NFAT response elements.

TCRα/β knockout in the TCR Knockout NFAT-Luciferase Reporter Jurkat cells was confirmed by genomic sequencing and by flow cytometry and expression of CD8 was confirmed by flow cytometry. The cell line has been functionally validated and does not respond to anti-CD3 agonist antibodies, as opposed to parental NFAT-Luciferase Reporter Jurkat cells (BPS Bioscience #60621).

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

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Synonyms

T-lymphocyte differentiation antigen T8/Leu-2, CD8a, MAL

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Product Data Gallery

Expression of CD8 in CD8+ TCR Knockout NFAT-Luciferase Reporter Jurkat cells.

Expression of transgenic TCR in Jurkat cells transduced with TCR lentiviruses

The Expression of Transgenic TCR in CD8+ TCR knockout NFAT Luciferase Reporter Cell Line confers responsiveness to anti-CD3 agonist, which induces NFAT-dependent luciferase activity.

T Cell Activation using T2 cells as APC.

Product Info

Storage and Usage

Citations

Host Cell Line

Jurkat (clone E6-1), human T lymphoblast, suspension

Supplied As

Each vial contains 2 x 10⁶ cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796)

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
Anti-CD3 Agonist Antibody	BPS Bioscience #71274
NY-ESO-1 Specific TCR Lentivirus (1G4)	BPS Bioscience #78675
MART-1 Specific TCR Lentivirus (DMF4)	BPS Bioscience #78678
T2 Cell Line	ATCC #CRL-1992
NY-ESO-1 (157-165) Peptide	BPS Bioscience #78758
MART-1 (Leu26-35, Leu27) Peptide	BPS Bioscience #78760
APC MHC I Dextramer (HLA-A*02:01 SLLMWITQV)	Immudex #WB03247
APC MHC I Dextramer (HLA-A*02:01 ELAGIGILTV)	Immudex #WB02162
Alexa Fluor 647™ Anti-human CD8	Biolegend #344725
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
96-well tissue culture plate, white, clear bottom

Genbank #

NM_001768.6

UniProt #

P01732

Mycoplasma Testing

The cell line has been screened to confirm the absence of Mycoplasma species.

Background

The TCR (T Cell Receptor) protein complex is found on the surface of T cells and is responsible for recognizing antigens bound to MHC (Major Histocompatibility Complex) molecules. Stimulation of the TCR results in activation of downstream NFAT (Nuclear factor of Activated T-cells) signaling. NFAT is a family of transcription factors that has an important function in immune responses, for example by inducing the expression of various cytokines (such as interleukin-2 to 4, and TNF-alpha) in T cells. NFAT is regulated by Ca²⁺ and the Ca^2+/calmodulin-dependent serine phosphatase calcineurin.

The TCR consists of a heterodimer of two different protein chains, of which the alpha (α) and beta (β) chains are the predominant chains. CRISPR/Cas9 genome editing was used to remove the TRAC (T-Cell Receptor Alpha Constant) and TRBC1 (T-Cell Receptor Beta Constant 1) regions of the α and β chains, resulting in loss of TCR .

Storage/Stability

Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.

Growth Media

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

Media Required for Cell Culture
Thaw Medium 2 (BPS Bioscience #60184): RPMI 1640 medium supplemented with 10% FBS, and 1% Penicillin/Streptomycin.

Growth Medium 2C (BPS Bioscience #79592): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 1 mg/ml of Geneticin and 100 µg/ml Hygromycin B.

Media Used in Functional Cellular Assay
Assay Medium 2D (BPS Bioscience #78755): RPMI 1640 medium supplemented with 1% FBS

Applications

Study transgenic TCR expression by introducing TCR alpha and beta chains.
Study the activation of transgenic TCR by cognate peptide(s) and MHC1-expressing APCs (Antigen Presenting Cells).

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.

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

Troubleshooting Guide
Visit cell-line-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Immunotherapy

Regulatory Status

Research Use Only

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CD8+ TCR Knockout NFAT-Luciferase Reporter Jurkat Cell Line

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