KC-6064

HEK293-IL1RAP-KO Cell Line

60065

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Background of HEK293-IL1RAP-KO 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-6064
Cell Line Name	HEK293-IL1RAP-KO Cell Line
NCBI/UniProt Accession Number	3556
Clone Number	2A1
Host Cell Line	HEK293
Description	Stable HEK293 cell line with IL1RAP gene knockout, No.2A1
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	70% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

HEK293-IL1RAP-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HEK293-IL1RAP-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of HEK293-IL1RAP-KO Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of HEK293(Left) and HEK293-IL1RAP-KO(Right) Cell Line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM+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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

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.
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.