HK-CRISPR-mEGFP-Nup358

The HK-CRISPR-mEGFP-Nup358 cell line is a genetically engineered derivative of HeLa Kyoto cells, known for their robustness and widespread use in scientific research. This cell line has been modified using CRISPR-Cas9 technology to express mEGFP (monomeric Enhanced Green Fluorescent Protein) tagged Nup358, a crucial component of the nuclear pore complex (NPC). Nup358, also known as RanBP2, plays a significant role in nucleocytoplasmic transport, mitotic spindle assembly, and other cellular processes. The mEGFP tag allows for the visualization of Nup358, facilitating real-time observation of its dynamics and interactions within the cell.

HeLa Kyoto cells, a subline of the original HeLa cells, are characterized by their adaptability and stable growth in culture. The CRISPR-Cas9 system in this cell line enables precise genomic editing, ensuring the mEGFP tag is accurately fused to the Nup358 protein without disrupting its function. This makes the HK-CRISPR-mEGFP-Nup358 cell line a valuable tool for studying the structural and functional aspects of the nuclear pore complex. Researchers can use this cell line to gain insights into the mechanisms governing nucleocytoplasmic transport and the role of Nup358 in cellular homeostasis and disease states, such as cancer and viral infections.

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.