KC-4149

Jurkat TCR CD4 KO 3C4 Cell Line

43780

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Background of Jurkat TCR CD4 KO 3C4 Cell Line

This gene encodes the CD4 membrane glycoprotein of T lymphocytes. The CD4 antigen acts as a coreceptor with the T-cell receptor on the T lymphocyte to recognize antigens displayed by an antigen presenting cell in the context of class II MHC molecules. The CD4 antigen is also a primary receptor for entry of the human immunodeficiency virus through interactions with the HIV Env gp120 subunit. This gene is expressed not only in T lymphocytes, but also in B cells, macrophages, granulocytes, as well as in various regions of the brain. The protein functions to initiate or augment the early phase of T-cell activation, and may function as an important mediator of indirect neuronal damage in infectious and immune-mediated diseases of the central nervous system. Multiple alternatively spliced transcript variants encoding different isoforms have been identified in this gene.

Specifications

Catalog Number	KC-4149
Cell Line Name	Jurkat TCR CD4 KO 3C4 Cell Line
Host Cell Line	Jurkat-TCR-KO-1B1 Cell Line
Description	Stable Jurkat-TCR-KO-1B1 clone with human CD4 gene knockout, No.3C4
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	N/A
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 26 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Jurkat-TCR-CD4-KO-3C4 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of TRAC knockout in Jurkat using PCR sequencing.

Figure 2: Characterization of TRBC knockout in Jurkat-TRAC-KO-2B1 using PCR sequencing.

Figure 3: Characterization of TRBC knockout in Jurkat-TRAC-KO-2B1 using RT-PCR sequencing.

Figure 4: Characterization of TCR knockout in Jurkat using FACS.

Figure 5: Characterization of CD4 knockout in Jurkat-TCR-KO-1B1 using PCR sequencing.

Figure 6: Characterization of CD4 knockout in Jurkat-TCR-KO-1B1 using RT-PCR sequencing.

Figure 7: Characterization of CD4 knockout in Jurkat-TCR-KO-1B1 using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

https://www.ncbi.nlm.nih.gov/gene/920