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HK-ZFN-AURKB-mEGFP genomic DNA - 5 microgram

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Cat# 300173GD5

Size : 5ug

Brand : CLS Cell Lines Service

HK-ZFN-AURKB-mEGFP genomic DNA - 5 microgram

HK-ZFN-AURKB-mEGFP Cells

HK-ZFN-AURKB-mEGFP Cells

HK-ZFN-AURKB-mEGFP Cells

HK-ZFN-AURKB-mEGFP Cells

HK-ZFN-AURKB-mEGFP Cells

Description	The HK-ZFN-AURKB-mEGFP cell line is a genetically engineered human cell model designed to express the AURKB (Aurora Kinase B) protein fused with mEGFP (monomeric Enhanced Green Fluorescent Protein) using Zinc Finger Nuclease (ZFN) technology. AURKB is a serine/threonine kinase that plays a crucial role in mitotic chromosome segregation, cytokinesis, and the regulation of the mitotic spindle checkpoint. The fusion with mEGFP allows for real-time visualization of AURKB activity and localization within the cell, facilitating detailed studies of its dynamic behavior during cell division. This cell line serves as a powerful tool for researchers investigating the molecular mechanisms of mitosis and the specific functions of AURKB. The incorporation of mEGFP enables fluorescence-based assays and live-cell imaging, providing insights into the spatiotemporal distribution of AURKB. The use of ZFN technology ensures precise genomic integration, maintaining the fidelity of AURKB expression. This model is particularly valuable in cancer research, where AURKB is often overexpressed and linked to tumorigenesis, making it a potential target for therapeutic interventions.
Organism	Human
Tissue	Endocervix
Disease	Adenocarcinoma

Age	30 years
Gender	Female
Ethnicity	African American
Morphology	Epithelial-like cells with mosaic stone shape
Growth properties	Adherent

Citation	HK-ZFN-AURKB-mEGFP (Cytion catalog number 300173)
Biosafety level	1
Depositor	Dr. J. Ellenberg, EMBL Heidelberg

Products	EGFP (Enhanced Green Fluorescent Protein)

Culture Medium	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)
Medium supplements	Supplement the medium with 10% FBS
Passaging solution	Accutase
Subculturing	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.
Split ratio	A ratio of 1:3 is recommended
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)
Handling of cryopreserved cultures	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. Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours. 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.

Sterility	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.