KC-5796

CHOK1-cyno-IL1RAP-Cell-Line

63935

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Background of CHOK1-cyno-IL1RAP-Cell-Line

The interleukin 1 receptor accessory protein (IL1RAP) gene, located on human chromosome 3 (191,714,585-191,858,537), encodes a critical co-receptor in IL-1 family signaling networks. It forms heteromeric complexes with ligand-binding receptors (e.g., ST2 for IL-33), facilitating adaptor protein recruitment (e.g., MyD88) to activate NF-κB and MAPK pathways, regulating immune responses and cytokine production. Alternative splicing generates membrane-bound and soluble isoforms; the soluble form modulates inflammation by enhancing ligand-decoy receptor binding. Overexpressed in hematologic malignancies (e.g., acute myeloid leukemia) and solid tumors (e.g., Ewing’s sarcoma), IL1RAP is a promising therapeutic target for CAR-T and antibody-based therapies.

Specifications

Catalog Number	KC-5796
Cell Line Name	CHOK1-cyno-IL1RAP-Cell-Line
NCBI/UniProt Accession Number	XM_005545409.5
Clone Number	5#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous cyno-IL1RAP gene.
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	70% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640+10%FBS+750μg/mL Hygromycin B
Selection Marker	Hygromycin B
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-cyno-IL1RAP-cell-line was generated using a lentiviral vector expressing the cyno-IL1RAP sequence.

Characterization

Figure 1: Characterization of cyno-IL1RAP overexpression in the CHOK1-cyno-IL1RAP stable clone using FACS.

Figure 2: Characterization of cyno-IL1RAP in the CHOK1-cyno-IL1RAP stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 750μg/mL Hygromycin B)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

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

References

1.Frenay J, Bellaye PS, Oudot A, Helbling A, Petitot C, Ferrand C, Collin B, Dias AMM. IL-1RAP, a Key Therapeutic Target in Cancer. Int J Mol Sci. 2022 Nov 29;23(23):14918. doi: 10.3390/ijms232314918. PMID: 36499246; PMCID: PMC9735758.
2.Weber A, Wasiliew P, Kracht M. Interleukin-1 (IL-1) pathway. Sci Signal. 2010 Jan 19;3(105):cm1. doi: 10.1126/scisignal.3105cm1. PMID: 20086235.

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.