联系我们
[gtranslate]
Contact

KC-4425

HCT116-WRN-C727S-KI-3A1 Cell Line

×
请在浏览器中启用JavaScript来完成此表单。
Datasheet
47786
Home » HCT116-WRN-C727S-KI-3A1 Cell Line

Background of HCT116-WRN-C727S-KI-3A1 Cell Line

The WRN gene, encoding the Werner syndrome RecQ-like helicase (WRN), is a member of the RecQ helicase family. This protein plays a crucial role in maintaining genomic stability through its involvement in DNA repair, replication, and telomere maintenance. Mutations in the WRN gene are responsible for Werner syndrome, a rare autosomal recessive disorder characterized by premature aging and an increased risk of cancer. Individuals with Werner syndrome exhibit symptoms such as early onset of atherosclerosis, type 2 diabetes, osteoporosis, and cataracts, typically developing these conditions in their 20s or 30s.

Specifications

Catalog NumberKC-4425
Cell Line NameHCT116-WRN-C727S-KI-3A1 Cell Line
Host Cell LineHCT116
DescriptionStable HCT116 clone expressing exogenous WRN gene bearing C727S mutations, No.3A1
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing MediumMcCoy‘s 5a+20% FBS+10% DMSO
Propagation MediumMcCoy‘s 5a+10% FBS
Selection MarkerN/A
MorphologyEpithelial
SubcultureSplit saturated culture 1:3-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative

Cell Line Generation

HCT116-WRN-C727S-3A1 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HCT116-WRN-C727S-KI-3A1 cell line stable clone using PCR sequencing.

Figure 2: Characterization of HCT116-WRN-C727S-KI-3A1 cell line stable clone using RT-PCR sequencing.

Figure 3. Characterization of dose-response curves for WRN inhibitors on HCT116 and HCT116-WRN-C727S-KI-3A1 cells.

Cell Resuscitation

  1. Prewarm culture medium (McCoy‘s 5a + 10% FBS)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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% McCoy‘s 5a + 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. Opresko, P. L., et al. (2014). The Werner syndrome helicase and exonuclease cooperate to resolve telomeric D loops in a manner regulated by TRF1 and TRF2.Molecular Cell, 54(6), 1163-1174. https://doi.org/10.1016/j.molcel.2014.05.018
  2. Monnat, R. J., Jr. (2010). Human RECQ helicases: roles and mechanisms.Nature Reviews Molecular Cell Biology, 11(10), 721-733. https://doi.org/10.1038/nrm2970
  3. Oshima, J., et al. (2017). Werner syndrome: a review on clinical features, molecular mechanisms, and potential therapies.Aging and Disease, 8(5), 624-644. https://doi.org/10.14336/AD.2017.0115

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.

Get a Quote

请在浏览器中启用JavaScript来完成此表单。