U2OS-CRISPR-SNAPf-SEH1 Cells

USD 800.00*

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).

Los precios no incluyen impuestos ni gastos de envío

cryovial

Número de producto: 300664

Ficha de datos de seguridad

Ficha del producto

Descripción	U2OS-CRISPR-SNAPf-SEH1 is a genome-edited human osteosarcoma cell line derived from U2OS cells in which the endogenous SEH1L (SEH1) gene has been modified using CRISPR/Cas9 technology to encode an in-frame SNAPf tag. SEH1 is a component of the Y-complex (also known as the NUP107-160 complex), a core structural module of the nuclear pore complex (NPC) that contributes to pore scaffold assembly and stability. By inserting the SNAPf coding sequence at the endogenous locus, the tagged SEH1 protein is expressed under native regulatory control, preserving physiological expression levels and minimizing perturbations to nuclear pore composition. The SNAPf tag is an engineered, fast-reacting variant of the SNAP-tag that covalently binds benzylguanine-conjugated substrates, allowing selective and stable fluorescent labeling in live or fixed cells. In U2OS-CRISPR-SNAPf-SEH1 cells, the fusion protein localizes to the nuclear envelope in a punctate pattern characteristic of NPC distribution. Because labeling occurs at endogenous protein levels, this system is well suited for quantitative fluorescence microscopy, super-resolution imaging, and single-particle tracking analyses aimed at dissecting NPC organization and stoichiometry. The flat morphology and large nuclei of U2OS cells further facilitate high-resolution visualization of nuclear envelope structures. SEH1 participates in NPC biogenesis and has also been implicated in kinetochore-associated processes during mitosis. Accordingly, this cell line provides a robust platform for investigating cell cycle-dependent NPC assembly and disassembly, spatial organization of the Y-complex within the pore scaffold, and potential dual roles of SEH1 at the nuclear envelope and mitotic kinetochores. U2OS-CRISPR-SNAPf-SEH1 enables mechanistic studies of nuclear pore architecture and dynamics under physiologically relevant expression conditions.
Organismo	Human
Tejido	Bone
Enfermedad	Osteosarcoma

Edad	15 years
Género	Female
Origen étnico	Caucasian
Morfología	Epithelial-like
Propiedades de crecimiento	Adherent

Referencia	U2OS-CRISPR-SNAPf-SEH1 (Cytion catalog number 300664)
Nivel de bioseguridad	1
NCBI_TaxID	9606
Depositante	The Ellenberg Lab (EMBL)
Estado de los OGM	GMO-S1: This human osteosarcoma cell line (U2OS-CRISPR-SNAPf-SEH1) contains a CRISPR-mediated SNAPf-SEH1 fusion that allows selective labeling of the SEH1 nucleoporin. The modification is stably present. This classification applies only within Germany and may differ elsewhere.

Expresión de proteínas	SEH1, SNAPf-tag

Medio de cultivo	McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a)
Suplementos	Supplement the medium with 10% FBS, 3.0 g/L Glucose, stable Glutamine, 2.0 mM Sodium pyruvate, 2.2 g/L NaHCO3, 1% NEAA
Reactivo de disociación	Accutase
Subcultivo	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.
Medio de congelación	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.
Descongelación y cultivo de células	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.
Atmósfera de incubación	37°C, 5% CO₂, humidified atmosphere.
Condiciones de envío	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.
Condiciones de almacenamiento	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.

Esterilidad	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.

U2OS-CRISPR-SNAPf-SEH1 Cells

Información general

Características

Datos normativos

Datos biomoleculares

Manejo

Control de calidad y análisis molecular