KC-5066

Jurkat-IKZF1-HiBiT-KI(+/-) Cell Line

61447

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Background of Jurkat-IKZF1-HiBiT-KI(+/-) Cell Line

IKAROS is part of the IKAROS protein family together with AIOLOS (IKZF3), HELIOS (IKZF2), EOS (IKZF4), and PEGASUS (IKZF5) [Citation4]. These comprise an important family of zinc finger transcription factors that regulate the differentiation, proliferation, and survival of lymphocytes. The IKZF1 gene encoding the IKAROS protein is located on the short arm of chromosome 7 on band 12.2. This gene contains eight exons and the protein is encoded by exons 2–8. The IKAROS protein is characterized by six zinc finger motifs. The four N-terminal zinc fingers, encoded by exons 4–6, mediate binding to DNA sequences throughout the genome. Exon 8 encodes the two zinc-fingers enabling the protein to homodimerize or heterodimerize with other transcription factors, including other members of the IKAROS family.

Specifications

Catalog Number	KC-5066
Cell Line Name	Jurkat-IKZF1-HiBiT-KI(+/-) Cell Line
Clone Number	1A3
Host Cell Line	Jurkat
Description	Stable Jurkat clone expressing IKZF1 with an endogenous C-terminal HiBiT tag
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	NA
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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

Jurkat-IKZF1-HiBiT-KI(+/-) cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Jurkat-IKZF1-HiBiT-KI(+/-) cell line stable clone using PCR sequencing.

Figure 2:Characterization of Jurkat-IKZF1-HiBiT-KI(+/-) cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of HiBiT degradation by IKZF1 Degrader.

Figure 4:Characterization of Jurkat-IKZF1-HiBiT-KI(+/-) Cell Line stable clone using Nano-Glo® HiBiT Lytic Detection System in the conditions of different cell number.

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% 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

Østergaard A, Boer JM, van Leeuwen FN, Pieters R, Den Boer ML. IKZF1 in acute lymphoblastic leukemia: the rise before the fall? Leuk Lymphoma. 2024 Dec;65(14):2077-2087. doi: 10.1080/10428194.2024.2396046. Epub 2024 Aug 29. PMID: 39210599.