KC-1405

HCT116-MTAP-KO Cell Line

22075

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Background of HCT116-MTAP-KO Cell Line

MTAP (methylthioadenosine phosphorylase) is encoded by the MTAP gene located on chromosome 9p21.3 and serves as a key metabolic enzyme. This chromosomal region often undergoes deletion in various cancers, including glioblastoma, mesothelioma, and non-small cell lung cancer, frequently co-occurring with CDKN2A/B gene deletions. The MTAP enzyme catalyzes the phosphorylation of 5\'-deoxy-5\'-methylthioadenosine (MTA), a byproduct of polyamine synthesis, to yield adenine and methylthioribose-1-phosphate. This reaction is essential for recycling purine precursors and maintaining cellular methionine levels, linking MTAP activity to nucleotide biosynthesis and epigenetic regulation mediated by S-adenosylmethionine (SAM).

Specifications

Catalog Number	KC-1405
Cell Line Name	HCT116-MTAP-KO Cell Line
Clone Number	11B1
Host Cell Line	HCT116
Description	Stable HCT116 clone with human MTAP gene knockout, No.11B1
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	70% McCoy's 5A+20% FBS+10% DMSO
Propagation Medium	McCoy's 5A+10% FBS
Selection Marker	NA
Morphology	Fibroblast
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

HCT116-MTAP-KO-11B1 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HCT116-MTAP-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of HCT116-MTAP-KO cell line stable clone using western blot.

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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% 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

Miao D, Li Y, Shen X, et al. MTAP deletion and immune checkpoint inhibitor resistance in solid tumors. J Clin Oncol. 2020 Mar 15;38(15_suppl):11546. doi:10.1200/JCO.2020.38.15_suppl.11546. PMID:32256891.
Vazquez F, Zhang R, Qin X, et al. Targeting MTAP-deficient tumors through metabolic rewiring. Cancer Discov. 2021 Aug;11(8):1972-1990. doi:10.1158/2159-8290.CD-20-1582. Epub 2021 Jun 14. PMID:34136785; PMCID:PMC8363878.

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.