KC-1888

293T-GPRC5A Cell Line

22993

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Background of 293T-GPRC5A Cell Line

GPRC5A, also known as RAI3, is a protein that encodes a member of the type 3 G protein-coupled receptor family.GPRC5A may be involved in interaction between retinoic acid and G protein signaling pathways. GPRC5A plays a critical role in development, cellular growth, and differentiation and it may play a role in embryonic development and epithelial cell differentiation. GPRC5A is dysregulated in many human cancers and in other diseases.

Specifications

Catalog Number	KC-1888
Cell Line Name	293T-GPRC5A Cell Line
NCBI/UniProt Accession Number	NM_003979.4
Host Cell Line	293T
Description	Stable HEK293T cell line expressing exogenous GPRC5A gene
Quantity	One vial of frozen cells (≥2×10⁶ cells)
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 + 1µg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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
In Vivo Validation	NA

Cell Line Generation

293T-GPRC5A cell line was generated using a lentiviral vector expressing the GPRC5A sequence.

Characterization

Figure 1:Characterization of 293T-GPRC5A cell line stable clone using FACS.

Figure 2: Characterization of 293T-GPRC5A cell line stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/ml Puromycin) 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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.Cheng Y, Lotan R (1998). "Molecular cloning and characterization of a novel retinoic acid-inducible gene that encodes a putative G protein-coupled receptor". J. Biol. Chem. 273 (52): 35008–15. doi:10.1074/jbc.273.52.35008. PMID 9857033.
2.Cafferata EG, Gonzalez-Guerrico AM, Pivetta OH, Santa-Coloma TA (1996). "Identification by differential display of a mRNA specifically induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in T84 human colon carcinoma cells". Cell. Mol. Biol. (Noisy-le-grand). 42 (5): 797–804. PMID 8832110. 4.Zhou H, Rigoutsos I (2014). "The emerging roles of GPRC5A in diseases". Oncoscience. 1 (12): 765–76. doi:10.18632/oncoscience.104. PMC 4303886. PMID 25621293.

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.