NRK-4xlambdaN22-3xmEGFP-M9 Cells

The NRK-4xlambdaN22-3xmEGFP-M9 cell line is a clonal stable cell line derived from normal rat kidney (NRK) cells through the transfection of a circular plasmid. This plasmid contains genetic constructs encoding four tandem repeats of lambda N22 RNA-binding sites and three tandem repeats of mEGFP (monomeric enhanced green fluorescent protein) tags fused with the M9 nuclear localization signal. Post-transfection, the cells underwent drug resistance selection to ensure the stability of the genetic modifications.

Approximately 50% of the cells in this clonal stable line express the fluorescent marker 4xλN22-3xmEGFP-M9, indicating successful incorporation of the plasmid. The expression of this marker allows for real-time visualization of intracellular processes, facilitated by the robust fluorescent signal of mEGFP. The M9 nuclear localization signal ensures that the expressed fusion proteins are transported to the nucleus, making this cell line particularly useful for studying nuclear-cytoplasmic transport, RNA dynamics, and gene expression regulation. However, it is important to note that this cell line exhibits some variegation, which refers to the variability in gene expression levels among individual cells within the population.

This NRK-4xlambdaN22-3xmEGFP-M9 cell line is valuable for researchers focusing on RNA-binding protein interactions, RNA metabolism, and the mechanisms underlying nuclear import and export. The presence of the mEGFP marker enables advanced imaging techniques such as confocal microscopy and live-cell imaging, providing detailed insights into the spatial and temporal dynamics of cellular components. Despite the variegation, the cell line remains a powerful tool for dissecting complex molecular pathways and understanding cellular functions at a deeper level.

Monolayer, adherent

Fibroblast-like cells with fusiform shape

DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)

Discard the old medium and wash the cells with PBS. Add a freshly prepared 0.025% trypsin/0.02% EDTA solution heated to 37 degrees Celsius and wait until the cells detach, which usually takes about 5 minutes. Neutralize the trypsin by adding fresh medium, then transfer the cell mixture to a tube and centrifuge. After centrifugation, remove the supernatant, resuspend the cell pellet in fresh culture medium, and transfer the suspension to new flasks. Incorporate G418 into the culture medium to achieve a final concentration of 0.5 mg/ml

CM-1 (Cytion catalog number 800100)

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.

Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.

A ratio of 1:3 to 1:4 is recommended

M9-His tag between BsrG1/HindIII, Neomycin, Phosphotransferase, CMV Promotor

Epidermal growth factor (EGF), multiplication stimulating activity (MSA)