| Field | Specification |
|---|---|
| Organism | |
| Applications | |
| Tissue | |
| Disease | Colorectal carcinoma |
| Age | |
| Sex | |
| Ethnicity | |
| Growth properties | |
| Biosafety level | |
| Product format | Frozen |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Overview
This product is a stable CRISPR knockout cell line in which ADRM1 (Adhesion Regulating Molecule 1, also known as RPN13) has been disrupted in the HCT116 human colorectal carcinoma background. The knockout was generated through CRISPR-Cas9–directed targeting of exon 4, replaced with a puromycin resistance cassette and pBluescriptII. This isogenic model supports investigation of proteasome biology, ubiquitin-mediated protein degradation, and cancer-related signaling. Cells grow as adherent, polygonal monolayers and are supplied frozen (BSL-II). An isogenic control cell line is available separately (Cat. No. T8240).
CRISPR Knockout Design
- Target gene: ADRM1 (Adhesion Regulating Molecule 1; also ADRM1/RPN13)
- Knockout strategy: CRISPR-Cas9–directed knock-in targeting exon 4; exon 4 was replaced with a puromycin resistance cassette and pBluescriptII backbone to enable selection of successfully transformed cells
- Selection marker: Puromycin resistance; 0.4 µg/ml puromycin used for selection (add only after first passage post-thaw)
- Parental cell line: HCT116 — human colorectal carcinoma; adherent, polygonal; derived from a 48-year-old Caucasian male donor
- Isogenic control: Available separately as Cat. No. T8240
Gene Background
ADRM1 (Adhesion Regulating Molecule 1), also referred to as RPN13 or ADRM1/RPN13, is a component of the 26S proteasome lid complex and functions as a ubiquitin receptor. It recruits the deubiquitinating enzyme UCHL5 (ubiquitin carboxyl-terminal hydrolase L5/UCH37) to the proteasome, facilitating substrate processing and ubiquitin chain editing. ADRM1 is known to undergo O-linked glycosylation, which may influence its interactions within the proteasome complex. Dysregulation of ADRM1 expression has been reported in several cancer types, including ovarian and colorectal carcinomas, where elevated expression has been associated with tumor progression. These characteristics make ADRM1 a subject of interest for cancer biology and drug target studies.
Cell Culture Specifications
- Recommended vessels
- PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422)
- Culture medium
- DMEM, High Glucose (TM500) + 10% FBS (do not heat-inactivate) + 1% Penicillin/Streptomycin Solution (G255)
- Culture conditions
- 37.0°C, 5% CO₂
- Selection
- 0.4 µg/ml Puromycin (G264); add only after first passage post-thaw
- Growth properties
- Adherent, polygonal
- Seeding density
- 30,000–50,000 cells/cm²
- Organism
- Human (H. sapiens)
- Tissue origin
- Colon (colorectal carcinoma)
- Biosafety level
- BSL-II
- Format
- Frozen; 1×10⁶ cells / 1.0 ml
Knockout is confirmed by the presence of frameshift-inducing INDELs, verified by Sanger Sequencing.
Research Applications
- Proteasome biology: Investigate the contribution of ADRM1 to 26S proteasome assembly, substrate recognition, and ubiquitin chain processing in a colorectal carcinoma model.
- Drug target validation: Assess the effect of proteasome inhibitors (e.g., bortezomib, carfilzomib) or ADRM1-targeted agents in ADRM1 KO vs. HCT116 parental and isogenic control (T8240).
- Ubiquitin pathway analysis: Profile global ubiquitination levels and specific substrate degradation using co-immunoprecipitation and mass spectrometry approaches.
- Carcinogenesis research: Study the role of ADRM1 dysregulation in colorectal cancer cell proliferation, survival, and drug sensitivity.
- Transcriptomic profiling: Identify downstream transcriptional consequences of ADRM1 loss using RNA-seq comparing KO, parental, and isogenic control lines.
Important Notes
- Selection drug timing: Add puromycin (0.4 µg/ml) to culture medium only after the first passage post-thaw to allow cells to recover from freeze-thaw conditions.
- Isogenic control: A matched control cell line (Cat. No. T8240) is available for paired experimental designs requiring isogenic comparison.
- Donor/source: HCT116 parental cells are derived from a 48-year-old Caucasian male with colorectal carcinoma.
- Viability warranty: Cell viability upon thaw is warranted for 30 days following shipment when handled according to abm's cell handling guidelines.
- Intended use: For laboratory research use only. Not intended for diagnostic, therapeutic, or clinical applications.
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
What is your warranty or return policy?
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
