EDITING RUNX2 KO IN B16 MELANOMA CELLS WITH CRISPR-CAS9 AS A POTENTIAL STRATEGY TO ENHANCE IN VIVO TUMOR RESPONSE TO THERAPEUTIC APPROACHES

Cutaneous melanoma (CM) is a highly heterogeneous and metastatic cancer with still limited effective therapeutic options. RUNX2, a transcription factor involved in key pathways such as apoptosis, epithelial–mesenchymal transition, and stemness, may play a role in melanoma progression and therapy resistance. This study describes the generation of RUNX2 knockout (KO) B16 melanoma cells using a CRISPR/Cas9 approach targeting exon 4. Following transfection, antibiotic selection, and clonal isolation, genomic editing was validated by PCR, Sanger sequencing, and Western blot. Seven edited clones were identified, including in-frame and heterozygous deletions, all of which lacked RUNX2 protein expression. This workflow provides a reliable model to investigate the role of RUNX2 in melanoma biology and its potential contribution to resistance mechanisms in preclinical models.