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.