KC-6308

Jurkat-IKZF2-C-HiBiT-KI-LgBiT Cell Line

64007

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Background of Jurkat-IKZF2-C-HiBiT-KI-LgBiT Cell Line

IKZF2 (IKAROS family zinc finger 2), also known as HELIOS, is a member of the Ikaros family of zinc finger transcription factors . It is predominantly expressed in hematopoietic cells, particularly regulatory T cells (Tregs) and natural killer (NK) cells, where it plays critical roles in lymphocyte development and immune regulation . IKZF2 contains an N-terminal DNA-binding domain with four zinc fingers and a C-terminal protein-binding domain mediating dimerization . Mechanistically, HELIOS maintains Treg stability and function by suppressing IL2 transcription . Germline dominant negative variants affecting the DNA-binding zinc fingers cause ICHAD syndrome (Immunodysregulation, Craniofacial anomalies, Hearing impairment, Athelia, Developmental delay), while loss-of-function variants primarily result in immunodeficiency and immune dysregulation without syndromic features . IKZF2 is also essential for outer hair cell maturation in the cochlea, with mutations leading to sensorineural hearing loss . Thus, IKZF2 serves as a critical regulator of both immune homeostasis and neurodevelopment.

Specifications

Catalog Number	KC-6308
Cell Line Name	Jurkat-IKZF2-C-HiBiT-KI-LgBiT Cell Line
Clone Number	1B2
Host Cell Line	Jurkat-LgBiT
Description	Stable Jurkat-LgBiT cell clone expressing endogenous C-terminal HiBiT tagged IKZF2
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+300μg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Lymphoblast
Subculture	Split saturated culture 1:3-1:6 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

Jurkat-IKZF2-C-HiBiT-KI-LgBiT cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Jurkat-IKZF2-C-HiBiT-KI-LgBiT cell line stable clone using PCR sequencing.

Figure 2: Characterization of Jurkat-IKZF2-C-HiBiT-KI-LgBiT cell line stable clone using RT-PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10%FBS+300μg/ml Hygromycin B)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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (RPMI1640+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

Mohajeri A, et al. (2023). "Dominant Negative Variants in IKZF2 Cause ICHAD Syndrome." Journal of Medical Genetics, 60(11):1092-1104 .
Vaseghi-Shanjani M, et al. (2025). "A Germline Heterozygous Dominant Negative IKZF2 Variant Causing Syndromic Primary Immune Regulatory Disorder and ICHAD." Journal of Clinical Immunology, 45:89 .

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.