Elimination of the Rassf9 Gene in the B16f0 Melanoma Line Through Crispr/Cas9

Melanoma is the cancer caused by the transformation of melanocytes, being the most lethal among skin cancers.It can be divided into four groups depending on the mutation present, with the most frequent changes in the B-Raf and Ras proteins.The Ras protein is associated with different signaling cascades, regulating processes such as proliferation, differentiation, morphology and apoptosis.The main effectors of Ras in cell death are members of the RASSF (Ras-association Family).RASSF9 has been implicated in keratinocyte differentiation and dermal homeostasis and, interestingly, its expression can be induced by sun exposure.To study the role of RASSF9 in melanomas, we will eliminate the expression of this gene in the B16F0 murine lineage, through the CRISPR/Cas9 system.After selecting a stable transfectant line (B16F0.R9KO), we will evaluate the impact of our manipulation on tumor growth in vitro and in vivo, as well as resistance to chemotherapy drugs.Finally, this lineage will serve as a tool for further studies on the role of RASSF9 in intracellular biochemical pathways and other aspects of tumor biology.