KC-4149

Jurkat TCR CD4 KO 3C4 Cell Line

43780

Home » Jurkat TCR CD4 KO 3C4 Cell Line

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	One vial 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

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.