HK-CRISPR-Nup93-mEGFP Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
Price range: €€€

1 104,00 CAD$*

Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 10⁶ cells for adherent lines or 5 × 10⁶ cells for suspension lines (refer to the batch CoA for details).

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cryovial

Numéro de produit : 300655

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Certificat d'analyse (CoA)

Description	The HK-CRISPR-Nup93-mEGFP cell line is derived from Hela Kyoto cells and engineered using CRISPR/Cas9 technology to express Nup93 fused with monomeric enhanced green fluorescent protein (mEGFP). This allows for real-time visualization of Nup93 within the nuclear envelope, aiding in the study of nuclear-cytoplasmic transport, nuclear pore complex assembly, and nuclear envelope integrity. Nup93 is crucial for maintaining nuclear pore complex architecture and function. The mEGFP tag enables tracking its dynamics and interactions, facilitating high-resolution imaging techniques like confocal microscopy. This cell line helps researchers understand gene regulation, nucleocytoplasmic trafficking, and cellular stress responses. The HK-CRISPR-Nup93-mEGFP cell line is a valuable resource for studying cell biology and nuclear pore complex-associated diseases, contributing to potential therapeutic strategies targeting nuclear transport pathways. It is particularly useful for exploring the nuclear envelope's role in cellular function and pathology.
Organisme	Human
Tissu	Endocervix
Maladie	Adenocarcinoma

Âge	30 years
Genre	Female
Origine ethnique	African American
Morphologie	Epithelial-like cells with mosaic stone shape
Propriétés de croissance	Adherent

Référence	HK-CRISPR-Nup93-mEGFP (Cytion catalog number 300655)
Niveau de biosécurité	1
NCBI_Numéro d'identification fiscale	9606
Déposant	The Ellenberg Lab (EMBL)
Statut OGM	GMO-S1: This HeLa Kyoto line contains an mEGFP knock-in at the endogenous Nup93 locus for nuclear pore structural studies. This classification applies only within Germany and may differ elsewhere.

Expression des protéines	Nup153, mEGFP-tag

Milieu de culture	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Suppléments	Supplement the medium with 10% FBS
Réactif de dissociation	Accutase
Repiquage	Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
Milieu de congélation	As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Décongélation et culture des cellules	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
Atmosphère d'incubation	37°C, 5% CO₂, humidified atmosphere.
Conditions d'expédition	Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
Conditions d'entreposage	For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Stérilité	Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Numéro de lot	Type de certificat	Date	Numéro de catalogue
300655-150123	Certificat d'analyse	23. May. 2025	300655

HK-CRISPR-Nup93-mEGFP Cells

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Certificat d'analyse (CoA)