NRAS mutant-A375 Isogenic Cell Line
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Cells contain cytomegalovirus (CMV) DNA sequences
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
NRASQ61K; BRAFV600E research, anti-cancer drug screening, BRAF drug resistant melanoma model, RAS-RAF kinase signaling pathway. Ideally the parental cell line (ATCC® CRL-1619™) is included as a control for drug-sensitive response in experiments with this cell line.
BRAF is a proto-oncogene encoding B-RAF, a serine/threonine kinase of the RAF family that acts downstream of RAS and upstream of MEK in the MAPK/ERK signaling pathway. Mutations in BRAF lead to excessive cellular proliferation, differentiation, and survival. BRAF V600E mutations are present in 50% of melanomas and although there are current BRAF inhibitors used as successful therapeutics, patients often become resistant to drugs several months following treatment. One mechanism of resistance to these inhibitors is caused by a secondary NRAS Q61K acquired mutation. CRL-1619IG-2 is an isogenic cell line created at ATCC by utilizing the CRISPR/Cas9 gene editing to generate a drug resistant NRAS Q61K mutation within the A375 melanoma cell line, which naturally habors the BRAF V600E mutation. The NRASQ61K mutation in CRL-1619IG-2 has been validated at the genomic, transcript, and protein bio-functional levels. CRL-1619IG-2 shows significant resistance to the BRAF inhibitors Dabrafineb and Vemurafenib when compared to its parental cell line in 2D and 3D culture systems. CRL-1619IG-2 can be a useful model to study the RAS–RAF–MEK–ERK–MAP kinase signaling pathway and to screen potential BRAF inhibitors and anti-cancer compounds for drug discovery and development.
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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Cormier JN, et al. Heterogeneous expression of melanoma-associated antigens and HLA-A2 in metastatic melanoma in vivo. Int. J. Cancer 75: 517-524, 1998. PubMed: 9466650
Sharma SD, et al. Melanotropic peptide-conjugated beads for microscopic visualization and characterization of melanoma melanotropin receptors. Proc. Natl. Acad. Sci. USA 93: 13715-13720, 1996. PubMed: 8943000
Gershwin ME, et al. Immunobiology of heterotransplanted human tumors in nude mice. J. Natl. Cancer Inst. 58: 1455-1463, 1977. PubMed: 857033
Giard DJ, et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51: 1417-1423, 1973. PubMed: 4357758