U-2 OS-CRISPR-NUP96-mEGFP clone no.195

The U-2 OS-CRISPR-NUP96-mEGFP is a genetically modified cell line derived from the human osteosarcoma U-2 OS parent line. This cell line incorporates a targeted insertion of the monomeric Enhanced Green Fluorescent Protein (mEGFP) tag at the NUP96 gene locus, achieved through the CRISPR-Cas9 gene-editing technology. NUP96, part of the nuclear pore complex, is essential for nuclear transport, and its fusion with mEGFP allows for real-time visualization of nuclear pore dynamics under fluorescent microscopy, providing valuable insights into nuclear transport mechanisms and nucleocytoplasmic trafficking.

This specific clone, numbered 195, has been selected for its stable expression of the NUP96-mEGFP fusion protein and maintains the typical characteristics of the U-2 OS lineage, including a robust cytoskeletal structure which is critical in studies related to cancer cell migration and metastasis. The application of CRISPR technology ensures precise gene editing, minimizing off-target effects which could compromise the integrity of the experimental outcomes. This makes U-2 OS-CRISPR-NUP96-mEGFP clone no.195 particularly useful for high-resolution imaging techniques and detailed cellular architecture studies, aiding in advanced research in cellular biology, cancer research, and nuclear transport phenomena.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

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.

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.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.