KC-6609

HEK293-IL1RAP-KO-rat-IL1RAP Cell Line

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Background of HEK293-IL1RAP-KO-rat-IL1RAP Cell Line

The interleukin 1 receptor accessory protein (IL1RAP) gene, located on human chromosome 3, encodes a critical co-receptor in IL-1 family signaling networks. It forms heteromeric complexes with ligand-binding receptors, facilitating adaptor protein recruitment 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 and solid tumors, IL1RAP is a promising therapeutic target for CAR-T and antibody-based therapies.

Specifications

Catalog Number	KC-6609
Cell Line Name	HEK293-IL1RAP-KO-rat-IL1RAP Cell Line
NCBI/UniProt Accession Number	NM_012968.1
Clone Number	1#
Host Cell Line	HEK293
Description	Stable HEK293-IL1RAP-KO clone expressing exogenous rat-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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS+150μg/mL Hygromycin B
Selection Marker	Hygromycin B
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-rat-IL1RAP-cell-line was generated using a lentiviral vector expressing the rat-IL1RAP sequence.

Characterization

Figure 1: Characterization of rat-IL1RAP overexpression in the HEK293-IL1RAP-KO-rat-IL1RAP stable clone using FACS.

Figure 2: Characterization of rat-IL1RAP in the HEK293-IL1RAP-KO-rat-IL1RAP stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM+10% FBS+150μg/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 1:4-1:8 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.

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.