KC-6093

293T-LRP5-LRP6-KO Cell Line

60220

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Background of 293T-LRP5-LRP6-KO Cell Line

The Low Density Lipoprotein Receptor Related Protein 6 (LRP6) gene encodes a key co receptor in the Wnt signaling pathway, which plays a critical role in embryonic development, cell proliferation, and tissue homeostasis. LRP6, along with LRP5, forms a family of single pass transmembrane proteins that interact with Wnt ligands to activate the canonical Wnt/β catenin signaling cascade.This pathway is essential for regulating gene expression involved in cell fate determination, organogenesis, and metabolic processes. Mutations in LRP6 have been linked to various human diseases, including osteoporosis, coronary artery disease, and cancer, highlighting its importance in both normal physiology and pathology.

Specifications

Catalog Number	KC-6093
Cell Line Name	293T-LRP5-LRP6-KO Cell Line
NCBI/UniProt Accession Number	4040
Clone Number	1A3
Host Cell Line	293T-LRP5-KO
Description	Stable 293T-LRP5-KO cell line with LRP6 gene knockout, No.1A3
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

293T-LRP5-LRP6-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 293T-LRP5-LRP6-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of 293T-LRP5-LRP6-KO Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of 293T-LRP5-KO(Left) and 293T-LRP5-LRP6-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

Clevers H, Nusse R. Wnt/β-catenin signaling and disease. Cell. 2012 Jun 8;149(6):1192-205. doi: 10.1016/j.cell.2012.05.012. PMID: 22682243.
MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell. 2009 Jul;17(1):9-26. doi: 10.1016/j.devcel.2009.06.016. PMID: 19619488; PMCID: PMC2861485.