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KC-3609

NCI-H1792-SHOC2-KO-1C4 Cell Line

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Home » NCI-H1792-SHOC2-KO-1C4 Cell Line

Background of NCI-H1792-SHOC2-KO-1C4 Cell Line

SHOC2 encodes a prototypical leucine-rich repeat protein that promotes downstream receptor tyrosine kinase (RTK)/RAS signaling and plays an important role in several cellular and developmental processes. Gain-of-function germ line mutations of SHOC2 drive the RASopathy Noonan-like syndrome, and SHOC2 mediates adaptive resistance to mitogen-activated protein kinase (MAPK) inhibitors. Similar to many scaffolding proteins, SHOC2 facilitates signal transduction by enabling proximal protein interactions and regulating the subcellular localization of its binding partners.

Specifications

Catalog NumberKC-3609
Cell Line NameNCI-H1792-SHOC2-KO-1C4 Cell Line
Host Cell LineNCI-H1792
DescriptionStable NCI-H1792 clone with human SHOC2 gene knockout, No.1C4
QuantityOne vial of frozen cells (≥2×106 cells)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing MediumRPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10% FBS
Selection MarkerNA
MorphologyEpithelial
SubcultureSplit saturated culture 1:2-1:5 every 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
In Vivo ValidationNA

Cell Line Generation

NCI-H1792-SHOC2-KO-1C4 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of SHOC2 knockout in NCI-H1792 using PCR sequencing.

Figure 2: Characterization of SHOC2 knockout in NCI-H1792 using RT-PCR sequencing.

Figure 3: Characterization of SHOC2 knockout in NCI-H1792 using western blot.

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640 + 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 1:2-1:5 every 3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% RPMI-1640 + 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. Kwon JJ, Hahn WC. A Leucine-Rich Repeat Protein Provides a SHOC2 the RAS Circuit: a Structure-Function Perspective. Mol Cell Biol. 2021 Mar 24;41(4):e00627-20. doi: 10.1128/MCB.00627-20. PMID: 33526449; PMCID: PMC8088128.

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.

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