KC-6116

HCT116-WRN-G729S-KI Cell Line

60250

Home » HCT116-WRN-G729S-KI Cell Line

Background of HCT116-WRN-G729S-KI Cell Line

Located on human chromosome 8p12, the WRN gene belongs to the RecQ helicase family, encoding a protein with helicase and exonuclease activities critical for DNA repair, telomere maintenance, and genome stability. Biallelic WRN mutations cause Werner syndrome, an adult-onset premature aging disorder with symptoms like premature graying and cataracts. The rare homozygous G729S missense mutation links to a distinct phenotype: progressive pulmonary failure, recurrent spontaneous pneumothorax, and interstitial lung disease (ILD) in adolescents/young adults. Unlike classic Werner syndrome, it lacks typical premature aging features. In silico analyses suggest G729S impairs WRN’s exonuclease activity or stability, disrupting DNA repair/telomere pathways, highlighting WRN’s pleiotropic roles.

Specifications

Catalog Number	KC-6116
Cell Line Name	HCT116-WRN-G729S-KI Cell Line
NCBI/UniProt Accession Number	7486
Clone Number	1A2
Host Cell Line	HCT116
Description	Stable HCT116 clone expressing endogenous WRN gene bearing G729S mutations, No.1A2
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	McCoy‘s 5a+20% FBS+10% DMSO
Propagation Medium	McCoy‘s 5a+10% FBS
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 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 27 hours
Mycoplasma Status	Negative

Cell Line Generation

HCT116-WRN-G729S-KI cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HCT116-WRN-G729S-KI Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of HCT116-WRN-G729S-KI Cell Line stable clone using RT PCR sequencing.

Figure 3: Characterization of Dose-response curves and IC50 values for HCT116 and HCT116-WRN-G729S-KI cells treated with VVD-214 and HRO761 over 8 days.

Cell Resuscitation

Prewarm culture medium (McCoy‘s 5a+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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (McCoy‘s 5a+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

Sezer A, Kayhan G, Gursoy TR, Eyuboglu TS, Percin FE. A homozygous missense variant in the WRN gene segregating in a family with progressive pulmonary failure with recurrent spontaneous pneumothorax and interstitial lung disease. Am J Med Genet A. 2023 Jan;191(1):220-227. doi: 10.1002/ajmg.a.62986. Epub 2022 Oct 10. PMID: 36214313.

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.